rowid,run.accession,experiment.accession,sample.accession,study.accession,bioproject,study.title,study.alias,study.type,study.abstract,study.attributes,study.PMIDs,sample.description,sample.title,sample.alias,sample.centername,sample.attributes,GEOsample.title,GEOsample.dataprocessing,GEOsample.source,GEOsample.treatmentprotocol,GEOsample.extractprotocol,GEOsample.growthprotocol,GEOsample.characteristics,GEOsample.accession,experiment.title,experiment.alias,experiment.library_name,experiment.design_description,experiment.library_construction_protocol,experiment.attributes,experiment.library_strategy,experiment.library_source,experiment.library_selection,experiment.library_layout,experiment.platform,experiment.instrument_model,experiment.spot_descriptor,experiment.study_ref,run.title,run.attributes,run.filename,run.semantic_name,run.total_bases,run.total_spots,run.alias,run.read_lengths,run.base_counts,run.r1_length,run.r2_length,run.r3_length,run.r4_length,run.Acount,run.Ccount,run.Gcount,run.Tcount,run.Ncount,run.experiment,run.pool_member,submission.accession,submission.srasource,submission.bioprojectsource,seqdetective.n_mates,seqdetective.mapping_rate.mate1,seqdetective.mapping_rate.mate2,seqdetective.nofeature_rate.mate1,seqdetective.nofeature_rate.mate2,seqdetective.sparsity.mate1,seqdetective.sparsity.mate2,seqdetective.pos_strand_rate.mate1,seqdetective.pos_strand_rate.mate2,seqdetective.readlen.mate1,seqdetective.readlen.mate2,seqdetective.judgement.mate1,seqdetective.judgement.mate2,seqdetective.judgement.reason,platform_family,instrument_generation,read_bias,selection_class,prep_kit,sc_or_bulk,tech_class,technology,tech_variant,submission.bioprojectsource.country,earliest_date,devstage_curation,devstage_curation_coarse,tissue_curation,tissue_curation_coarse 75652,SRR24758212,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_A01.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,196502541.0,3852991.0,GSM7432137 r1,0:51,A:49165276;C:47981089;G:48399038;T:50940008;N:17130,51,,,,49165276,47981089,48399038,50940008,17130,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.72705,,0.06867,,0.88552,,0.54207,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75653,SRR24758213,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_A10.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,57801819.0,1133369.0,GSM7432137 r10,0:51,A:15530697;C:12658068;G:12843873;T:16764026;N:5155,51,,,,15530697,12658068,12843873,16764026,5155,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.63663,,0.42833,,0.92111,,0.57817,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75654,SRR24758214,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_A11.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,72272559.0,1417109.0,GSM7432137 r11,0:51,A:18233220;C:16632293;G:17042518;T:20358144;N:6384,51,,,,18233220,16632293,17042518,20358144,6384,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.67945,,0.12353,,0.94095,,0.48607,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75655,SRR24758215,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_A12.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,187811019.0,3682569.0,GSM7432137 r12,0:51,A:49325726;C:43713702;G:44241940;T:50513058;N:16593,51,,,,49325726,43713702,44241940,50513058,16593,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.84473,,0.12213,,0.90666,,0.56528,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75656,SRR24758216,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_B01.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,24050070.0,471570.0,GSM7432137 r13,0:51,A:5907474;C:6114990;G:5982982;T:6042494;N:2130,51,,,,5907474,6114990,5982982,6042494,2130,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.47163,,0.30199,,0.93933,,0.53872,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75657,SRR24758217,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_B02.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,275774544.0,5407344.0,GSM7432137 r14,0:51,A:69654405;C:67726837;G:68298856;T:70070552;N:23894,51,,,,69654405,67726837,68298856,70070552,23894,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.84442,,0.07023,,0.88923,,0.53422,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75658,SRR24758218,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_B03.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,226648335.0,4444085.0,GSM7432137 r15,0:51,A:57531232;C:54256013;G:54956617;T:59884654;N:19819,51,,,,57531232,54256013,54956617,59884654,19819,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.80693,,0.10694,,0.9179,,0.52785,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75659,SRR24758219,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_B04.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,247590159.0,4854709.0,GSM7432137 r16,0:51,A:64151909;C:59176102;G:59606872;T:64633924;N:21352,51,,,,64151909,59176102,59606872,64633924,21352,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.85951,,0.0727,,0.88598,,0.52101,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75660,SRR24758220,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_B05.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,236963799.0,4646349.0,GSM7432137 r17,0:51,A:61256843;C:56596118;G:57122766;T:61967590;N:20482,51,,,,61256843,56596118,57122766,61967590,20482,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.86507,,0.0857,,0.89556,,0.38852,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75661,SRR24758221,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_B06.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,174345642.0,3418542.0,GSM7432137 r18,0:51,A:43218316;C:42711053;G:43771526;T:44629449;N:15298,51,,,,43218316,42711053,43771526,44629449,15298,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.85865,,0.02991,,0.91167,,0.40975,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75662,SRR24758222,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_B07.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,310253298.0,6083398.0,GSM7432137 r19,0:51,A:79341090;C:74303124;G:75662036;T:80919848;N:27200,51,,,,79341090,74303124,75662036,80919848,27200,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.87708,,0.0717,,0.89727,,0.46207,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75663,SRR24758223,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_A02.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,566253.0,11103.0,GSM7432137 r2,0:51,A:136988;C:128378;G:125521;T:175317;N:49,51,,,,136988,128378,125521,175317,49,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.67766,,0.11307,,0.95172,,0.5382,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75664,SRR24758224,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_B08.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,346341.0,6791.0,GSM7432137 r20,0:51,A:84780;C:84123;G:83720;T:93689;N:29,51,,,,84780,84123,83720,93689,29,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.63515,,0.09273,,0.96802,,0.52162,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75665,SRR24758225,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_B09.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,377961.0,7411.0,GSM7432137 r21,0:51,A:95293;C:90257;G:87563;T:104807;N:41,51,,,,95293,90257,87563,104807,41,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.69105,,0.08086,,0.96455,,0.53115,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75666,SRR24758226,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_B10.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,2250936.0,44136.0,GSM7432137 r22,0:51,A:539924;C:650712;G:518629;T:541473;N:198,51,,,,539924,650712,518629,541473,198,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.27445,,0.04803,,0.95276,,0.53885,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75667,SRR24758227,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_B11.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,207937659.0,4077209.0,GSM7432137 r23,0:51,A:52800304;C:49954044;G:50494891;T:54670231;N:18189,51,,,,52800304,49954044,50494891,54670231,18189,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.83348,,0.06056,,0.87643,,0.52466,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75668,SRR24758228,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_B12.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,116941317.0,2292967.0,GSM7432137 r24,0:51,A:31454818;C:25883611;G:26279868;T:33312749;N:10271,51,,,,31454818,25883611,26279868,33312749,10271,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.76622,,0.21094,,0.92413,,0.55894,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75669,SRR24758229,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_C01.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,539172.0,10572.0,GSM7432137 r25,0:51,A:134771;C:127049;G:127559;T:149745;N:48,51,,,,134771,127049,127559,149745,48,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.74289,,0.09475,,0.94769,,0.55262,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75670,SRR24758230,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_C02.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,249154176.0,4885376.0,GSM7432137 r26,0:51,A:62840979;C:61240493;G:61835852;T:63215177;N:21675,51,,,,62840979,61240493,61835852,63215177,21675,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.89326,,0.07803,,0.84967,,0.51569,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75671,SRR24758231,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_C03.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,641019.0,12569.0,GSM7432137 r27,0:51,A:175739;C:156164;G:141289;T:167772;N:55,51,,,,175739,156164,141289,167772,55,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.50169,,0.06287,,0.95611,,0.57318,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75672,SRR24758232,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_C04.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,223399839.0,4380389.0,GSM7432137 r28,0:51,A:58104465;C:53189406;G:53552668;T:58534071;N:19229,51,,,,58104465,53189406,53552668,58534071,19229,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.84775,,0.07316,,0.89883,,0.53245,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75673,SRR24758233,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_C05.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,2148528.0,42128.0,GSM7432137 r29,0:51,A:456160;C:713081;G:433891;T:545183;N:213,51,,,,456160,713081,433891,545183,213,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.31226,,0.04264,,0.94718,,0.54464,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75674,SRR24758234,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_A03.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,182987337.0,3587987.0,GSM7432137 r3,0:51,A:48471316;C:41287723;G:41835653;T:51376450;N:16195,51,,,,48471316,41287723,41835653,51376450,16195,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.79625,,0.12945,,0.87042,,0.54466,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75675,SRR24758235,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_C06.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,277189131.0,5435081.0,GSM7432137 r30,0:51,A:72722594;C:65268944;G:65970657;T:73202676;N:24260,51,,,,72722594,65268944,65970657,73202676,24260,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.85793,,0.09135,,0.89428,,0.53659,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75676,SRR24758236,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_C07.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,51855423.0,1016773.0,GSM7432137 r31,0:51,A:12581543;C:12652880;G:12719547;T:13896926;N:4527,51,,,,12581543,12652880,12719547,13896926,4527,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.64717,,0.1549,,0.93042,,0.59143,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75677,SRR24758237,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_C08.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,196417422.0,3851322.0,GSM7432137 r32,0:51,A:49916233;C:47717459;G:48374054;T:50392337;N:17339,51,,,,49916233,47717459,48374054,50392337,17339,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.75845,,0.10284,,0.91001,,0.55748,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75678,SRR24758238,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_C09.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,278052.0,5452.0,GSM7432137 r33,0:51,A:70438;C:67316;G:65222;T:75057;N:19,51,,,,70438,67316,65222,75057,19,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.70448,,0.07804,,0.96885,,0.56873,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75679,SRR24758239,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_C10.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,231233694.0,4533994.0,GSM7432137 r34,0:51,A:59674130;C:55503966;G:55963671;T:60071740;N:20187,51,,,,59674130,55503966,55963671,60071740,20187,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.88277,,0.07596,,0.85867,,0.55683,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75680,SRR24758240,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_C11.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,313956.0,6156.0,GSM7432137 r35,0:51,A:77901;C:74833;G:75253;T:85941;N:28,51,,,,77901,74833,75253,85941,28,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.64898,,0.08603,,0.96727,,0.56391,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75681,SRR24758241,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_C12.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,146542482.0,2873382.0,GSM7432137 r36,0:51,A:36685659;C:35025753;G:35662355;T:39155620;N:13095,51,,,,36685659,35025753,35662355,39155620,13095,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.8478,,0.07133,,0.88692,,0.56982,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75682,SRR24758242,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_D01.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,35044293.0,687143.0,GSM7432137 r37,0:51,A:8999254;C:8265360;G:8325013;T:9451693;N:2973,51,,,,8999254,8265360,8325013,9451693,2973,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.82477,,0.10365,,0.88185,,0.52857,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75683,SRR24758243,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_D02.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,10634316.0,208516.0,GSM7432137 r38,0:51,A:2725984;C:2435586;G:2465834;T:3005982;N:930,51,,,,2725984,2435586,2465834,3005982,930,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.81597,,0.16281,,0.87801,,0.57617,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75684,SRR24758244,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_D03.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,2658273.0,52123.0,GSM7432137 r39,0:51,A:638329;C:571678;G:578971;T:869067;N:228,51,,,,638329,571678,578971,869067,228,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.52947,,0.29121,,0.95526,,0.58642,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75685,SRR24758245,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_A04.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,197303394.0,3868694.0,GSM7432137 r4,0:51,A:50366781;C:47162624;G:47703993;T:52052535;N:17461,51,,,,50366781,47162624,47703993,52052535,17461,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.82032,,0.11001,,0.88619,,0.5478,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75686,SRR24758246,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_D04.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,14486397.0,284047.0,GSM7432137 r40,0:51,A:3735379;C:3368111;G:3417284;T:3964349;N:1274,51,,,,3735379,3368111,3417284,3964349,1274,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.81858,,0.10951,,0.88268,,0.54026,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75687,SRR24758247,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_D05.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,8936220.0,175220.0,GSM7432137 r41,0:51,A:2282631;C:2134511;G:2030647;T:2487667;N:764,51,,,,2282631,2134511,2030647,2487667,764,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.75044,,0.11145,,0.89879,,0.53216,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75688,SRR24758248,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_D06.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,17369223.0,340573.0,GSM7432137 r42,0:51,A:4545909;C:3952126;G:3972824;T:4896864;N:1500,51,,,,4545909,3952126,3972824,4896864,1500,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.80665,,0.13491,,0.89365,,0.5573,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75689,SRR24758249,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_D07.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,12178494.0,238794.0,GSM7432137 r43,0:51,A:3171768;C:2801174;G:2833782;T:3370712;N:1058,51,,,,3171768,2801174,2833782,3370712,1058,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.82506,,0.12982,,0.88245,,0.55146,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75690,SRR24758250,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_D08.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,13139079.0,257629.0,GSM7432137 r44,0:51,A:3368569;C:3043121;G:3093441;T:3632833;N:1115,51,,,,3368569,3043121,3093441,3632833,1115,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.84198,,0.13445,,0.90072,,0.5132,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75691,SRR24758251,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_D09.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,119040987.0,2334137.0,GSM7432137 r45,0:51,A:29695311;C:29202858;G:28762624;T:31369668;N:10526,51,,,,29695311,29202858,28762624,31369668,10526,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.88303,,0.0381,,0.91346,,0.59593,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75692,SRR24758252,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_D10.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,69416202.0,1361102.0,GSM7432137 r46,0:51,A:17390024;C:16946151;G:16559148;T:18514874;N:6005,51,,,,17390024,16946151,16559148,18514874,6005,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.86765,,0.03915,,0.91725,,0.63104,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75693,SRR24758253,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_D11.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,135864357.0,2664007.0,GSM7432137 r47,0:51,A:34526461;C:32359370;G:32701851;T:36264864;N:11811,51,,,,34526461,32359370,32701851,36264864,11811,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.85381,,0.07318,,0.87819,,0.54411,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75694,SRR24758254,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_D12.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,95575224.0,1874024.0,GSM7432137 r48,0:51,A:23888425;C:22595682;G:22919532;T:26163304;N:8281,51,,,,23888425,22595682,22919532,26163304,8281,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.79959,,0.12472,,0.88984,,0.57565,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75695,SRR24758255,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_E01.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,19084506.0,374206.0,GSM7432137 r49,0:51,A:4882904;C:4489652;G:4539293;T:5171086;N:1571,51,,,,4882904,4489652,4539293,5171086,1571,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.84491,,0.11267,,0.86927,,0.55631,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75696,SRR24758256,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_A05.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,153879036.0,3017236.0,GSM7432137 r5,0:51,A:39921479;C:36689938;G:36788395;T:40465732;N:13492,51,,,,39921479,36689938,36788395,40465732,13492,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.7994,,0.08355,,0.90522,,0.55633,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75697,SRR24758257,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_E02.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,26012550.0,510050.0,GSM7432137 r50,0:51,A:6739429;C:6056227;G:6086495;T:7128235;N:2164,51,,,,6739429,6056227,6086495,7128235,2164,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.83144,,0.1168,,0.8994,,0.52144,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75698,SRR24758258,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_E03.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,20283975.0,397725.0,GSM7432137 r51,0:51,A:5328019;C:4550331;G:4622714;T:5781195;N:1716,51,,,,5328019,4550331,4622714,5781195,1716,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.80043,,0.15348,,0.89643,,0.56281,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75699,SRR24758259,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_E04.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,2371857.0,46507.0,GSM7432137 r52,0:51,A:550735;C:548984;G:549060;T:722882;N:196,51,,,,550735,548984,549060,722882,196,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.34804,,0.23018,,0.97317,,0.49792,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75700,SRR24758260,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_E05.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,73613655.0,1443405.0,GSM7432137 r53,0:51,A:19134539;C:17308356;G:17281690;T:19882636;N:6434,51,,,,19134539,17308356,17281690,19882636,6434,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.8529,,0.08981,,0.89089,,0.50082,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75701,SRR24758261,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_E06.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,140298501.0,2750951.0,GSM7432137 r54,0:51,A:37003106;C:31738463;G:32324388;T:39220438;N:12106,51,,,,37003106,31738463,32324388,39220438,12106,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.85923,,0.11225,,0.88749,,0.45327,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75702,SRR24758262,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_E07.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,10771812.0,211212.0,GSM7432137 r55,0:51,A:2787721;C:2414278;G:2433404;T:3135476;N:933,51,,,,2787721,2414278,2433404,3135476,933,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.74729,,0.26126,,0.90707,,0.60814,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75703,SRR24758263,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_E08.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,20619708.0,404308.0,GSM7432137 r56,0:51,A:5331022;C:4777482;G:4809683;T:5699645;N:1876,51,,,,5331022,4777482,4809683,5699645,1876,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.76279,,0.14612,,0.90642,,0.58917,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75704,SRR24758264,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_E09.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,78138528.0,1532128.0,GSM7432137 r57,0:51,A:20291125;C:18267717;G:18451681;T:21121305;N:6700,51,,,,20291125,18267717,18451681,21121305,6700,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.86363,,0.09107,,0.89079,,0.51301,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75705,SRR24758265,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_E10.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,21589371.0,423321.0,GSM7432137 r58,0:51,A:5608520;C:4960262;G:4972805;T:6045881;N:1903,51,,,,5608520,4960262,4972805,6045881,1903,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.75496,,0.19769,,0.91194,,0.58205,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75706,SRR24758266,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_E11.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,7191510.0,141010.0,GSM7432137 r59,0:51,A:1607407;C:1664538;G:1655218;T:2263725;N:622,51,,,,1607407,1664538,1655218,2263725,622,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.27436,,0.16965,,0.97952,,0.53123,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75707,SRR24758267,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_A06.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,253757436.0,4975636.0,GSM7432137 r6,0:51,A:68746775;C:56465647;G:57533044;T:70990113;N:21857,51,,,,68746775,56465647,57533044,70990113,21857,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.86008,,0.10085,,0.94531,,0.43969,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75708,SRR24758268,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_E12.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,5967918.0,117018.0,GSM7432137 r60,0:51,A:1461664;C:1447795;G:1418933;T:1639051;N:475,51,,,,1461664,1447795,1418933,1639051,475,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.46793,,0.30628,,0.96662,,0.57737,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75709,SRR24758269,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_F01.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,54315.0,1065.0,GSM7432137 r61,0:51,A:13512;C:13799;G:13311;T:13689;N:4,51,,,,13512,13799,13311,13689,4,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.36191,,0.04857,,0.99492,,0.51552,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75710,SRR24758270,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_F02.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,120666.0,2366.0,GSM7432137 r62,0:51,A:30537;C:30751;G:27943;T:31426;N:9,51,,,,30537,30751,27943,31426,9,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.27847,,0.03283,,0.9919,,0.53546,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75711,SRR24758271,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_F03.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,80988.0,1588.0,GSM7432137 r63,0:51,A:22688;C:19856;G:18270;T:20166;N:8,51,,,,22688,19856,18270,20166,8,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.3473,,0.0527,,0.99391,,0.56074,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75712,SRR24758272,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_F04.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,431052.0,8452.0,GSM7432137 r64,0:51,A:91519;C:145281;G:87345;T:106862;N:45,51,,,,91519,145281,87345,106862,45,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.12561,,0.01174,,0.98802,,0.56926,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75713,SRR24758273,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_F05.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,1566822.0,30722.0,GSM7432137 r65,0:51,A:318939;C:552285;G:305938;T:389538;N:122,51,,,,318939,552285,305938,389538,122,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.10152,,0.01159,,0.98315,,0.513,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75714,SRR24758274,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_F06.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,66963.0,1313.0,GSM7432137 r66,0:51,A:17405;C:15737;G:15893;T:17919;N:9,51,,,,17405,15737,15893,17919,9,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.56077,,0.06615,,0.99196,,0.53785,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75715,SRR24758275,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_F07.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,64362.0,1262.0,GSM7432137 r67,0:51,A:14722;C:18342;G:14900;T:16394;N:4,51,,,,14722,18342,14900,16394,4,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.28836,,0.03453,,0.99614,,0.48717,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75716,SRR24758276,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_F08.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,32538.0,638.0,GSM7432137 r68,0:51,A:7742;C:8639;G:8538;T:7619;N:0,51,,,,7742,8639,8538,7619,0,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.19427,,0.03821,,0.99837,,0.52577,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75717,SRR24758277,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_F09.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,44880.0,880.0,GSM7432137 r69,0:51,A:10965;C:12433;G:10552;T:10923;N:7,51,,,,10965,12433,10552,10923,7,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.1857,,0.00922,,0.99868,,0.63815,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75718,SRR24758278,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_A07.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,438753.0,8603.0,GSM7432137 r7,0:51,A:107172;C:102802;G:100333;T:128413;N:33,51,,,,107172,102802,100333,128413,33,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.64165,,0.11997,,0.96418,,0.53578,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75719,SRR24758279,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_F10.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,1604715.0,31465.0,GSM7432137 r70,0:51,A:382053;C:482340;G:369584;T:370571;N:167,51,,,,382053,482340,369584,370571,167,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.09206,,0.01269,,0.9849,,0.57314,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75720,SRR24758280,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_F11.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,44931.0,881.0,GSM7432137 r71,0:51,A:11278;C:11046;G:10935;T:11667;N:5,51,,,,11278,11046,10935,11667,5,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.38991,,0.04931,,0.99582,,0.54827,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75721,SRR24758281,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_F12.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,54264.0,1064.0,GSM7432137 r72,0:51,A:13307;C:13960;G:12645;T:14346;N:6,51,,,,13307,13960,12645,14346,6,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.36477,,0.04857,,0.99515,,0.53703,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75722,SRR24758282,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_G01.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,223305897.0,4378547.0,GSM7432137 r73,0:51,A:56472139;C:53895417;G:54600016;T:58318632;N:19693,51,,,,56472139,53895417,54600016,58318632,19693,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.87754,,0.05837,,0.87771,,0.53057,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75723,SRR24758283,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_G02.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,242535702.0,4755602.0,GSM7432137 r74,0:51,A:60915597;C:59403540;G:60068417;T:62127178;N:20970,51,,,,60915597,59403540,60068417,62127178,20970,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.86621,,0.07791,,0.8677,,0.51049,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75724,SRR24758284,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_G03.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,212319222.0,4163122.0,GSM7432137 r75,0:51,A:54590684;C:50021453;G:50754851;T:56933780;N:18454,51,,,,54590684,50021453,50754851,56933780,18454,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.84687,,0.09197,,0.85259,,0.54054,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75725,SRR24758285,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_G04.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,208563990.0,4089490.0,GSM7432137 r76,0:51,A:53810659;C:49124991;G:49760146;T:55850022;N:18172,51,,,,53810659,49124991,49760146,55850022,18172,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.86502,,0.07038,,0.89092,,0.51629,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75726,SRR24758286,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_G05.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,222731739.0,4367289.0,GSM7432137 r77,0:51,A:56305313;C:54275918;G:54923070;T:57208464;N:18974,51,,,,56305313,54275918,54923070,57208464,18974,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.88652,,0.08367,,0.91654,,0.52511,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75727,SRR24758287,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_G06.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,529074.0,10374.0,GSM7432137 r78,0:51,A:129710;C:118983;G:121454;T:158896;N:31,51,,,,129710,118983,121454,158896,31,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.78205,,0.09486,,0.94868,,0.53186,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75728,SRR24758288,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_G07.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,16639566.0,326266.0,GSM7432137 r79,0:51,A:4136203;C:3917524;G:3860992;T:4723371;N:1476,51,,,,4136203,3917524,3860992,4723371,1476,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.83887,,0.09882,,0.92805,,0.59904,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75729,SRR24758289,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_A08.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,240618.0,4718.0,GSM7432137 r8,0:51,A:59927;C:55596;G:55853;T:69217;N:25,51,,,,59927,55596,55853,69217,25,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.61262,,0.12067,,0.97628,,0.53345,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75730,SRR24758290,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_G08.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,5564355.0,109105.0,GSM7432137 r80,0:51,A:1381661;C:1242035;G:1272174;T:1668047;N:438,51,,,,1381661,1242035,1272174,1668047,438,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.63091,,0.21785,,0.94132,,0.60924,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75731,SRR24758291,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_G09.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,9586878.0,187978.0,GSM7432137 r81,0:51,A:2479844;C:2172852;G:2222721;T:2710614;N:847,51,,,,2479844,2172852,2222721,2710614,847,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.82846,,0.1558,,0.89832,,0.5542,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75732,SRR24758292,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_G10.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,3365643.0,65993.0,GSM7432137 r82,0:51,A:798368;C:817432;G:806743;T:942805;N:295,51,,,,798368,817432,806743,942805,295,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.5385,,0.21274,,0.9544,,0.54539,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75733,SRR24758293,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_G11.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,4267323.0,83673.0,GSM7432137 r83,0:51,A:979559;C:1009380;G:1026556;T:1251434;N:394,51,,,,979559,1009380,1026556,1251434,394,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.32148,,0.25471,,0.96694,,0.54686,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75734,SRR24758294,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_G12.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,326298.0,6398.0,GSM7432137 r84,0:51,A:79720;C:78815;G:72411;T:95325;N:27,51,,,,79720,78815,72411,95325,27,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.62417,,0.09728,,0.9694,,0.56033,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75735,SRR24758295,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_H01.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,216877194.0,4252494.0,GSM7432137 r85,0:51,A:56096013;C:51068217;G:51596770;T:58097343;N:18851,51,,,,56096013,51068217,51596770,58097343,18851,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.82143,,0.10496,,0.89136,,0.54096,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75736,SRR24758296,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_H02.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,136640832.0,2679232.0,GSM7432137 r86,0:51,A:35885347;C:31385131;G:31657485;T:37700867;N:12002,51,,,,35885347,31385131,31657485,37700867,12002,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.7943,,0.14644,,0.9096,,0.57047,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75737,SRR24758297,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_H03.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,110655567.0,2169717.0,GSM7432137 r87,0:51,A:28864485;C:25160123;G:25604256;T:31017030;N:9673,51,,,,28864485,25160123,25604256,31017030,9673,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.7539,,0.1665,,0.90583,,0.54872,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75738,SRR24758298,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_H04.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,174712689.0,3425739.0,GSM7432137 r88,0:51,A:45561475;C:40736085;G:41075312;T:47324618;N:15199,51,,,,45561475,40736085,41075312,47324618,15199,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.83159,,0.09854,,0.87775,,0.52531,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75739,SRR24758299,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_H05.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,96307635.0,1888385.0,GSM7432137 r89,0:51,A:24570236;C:22312990;G:22574766;T:26841312;N:8331,51,,,,24570236,22312990,22574766,26841312,8331,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.78114,,0.20512,,0.88921,,0.61966,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75740,SRR24758300,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_A09.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,372147.0,7297.0,GSM7432137 r9,0:51,A:94549;C:92083;G:85469;T:100025;N:21,51,,,,94549,92083,85469,100025,21,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.58496,,0.094,,0.96804,,0.53823,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75741,SRR24758301,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_H06.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,255022797.0,5000447.0,GSM7432137 r90,0:51,A:66998458;C:58633157;G:59331325;T:70037873;N:21984,51,,,,66998458,58633157,59331325,70037873,21984,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.87604,,0.11589,,0.80277,,0.54722,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75742,SRR24758302,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_H07.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,78502005.0,1539255.0,GSM7432137 r91,0:51,A:20280221;C:18075885;G:18344442;T:21794509;N:6948,51,,,,20280221,18075885,18344442,21794509,6948,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.759,,0.16886,,0.92995,,0.57929,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75743,SRR24758303,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_H08.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,226950.0,4450.0,GSM7432137 r92,0:51,A:55730;C:52121;G:51716;T:67367;N:16,51,,,,55730,52121,51716,67367,16,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.65257,,0.1349,,0.97658,,0.57436,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75744,SRR24758304,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_H09.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,266883.0,5233.0,GSM7432137 r93,0:51,A:67933;C:63056;G:60189;T:75686;N:19,51,,,,67933,63056,60189,75686,19,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.60962,,0.10336,,0.97484,,0.59646,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75745,SRR24758305,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_H10.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,34504917.0,676567.0,GSM7432137 r94,0:51,A:8978757;C:7716865;G:7812277;T:9994055;N:2963,51,,,,8978757,7716865,7812277,9994055,2963,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.69725,,0.18041,,0.92547,,0.47385,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75746,SRR24758306,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_H11.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,254847.0,4997.0,GSM7432137 r95,0:51,A:62591;C:57094;G:57579;T:77558;N:25,51,,,,62591,57094,57579,77558,25,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.65802,,0.12628,,0.97348,,0.56099,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75747,SRR24758307,SRX20534082,SRS17842609,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep2,GSM7432137,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep2,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432137,GSM7432137: Photoconverted brain endothelial cells rep2; Danio rerio; RNA Seq,GSM7432137 r1,GSM7432137,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049751_H12.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,319056.0,6256.0,GSM7432137 r96,0:51,A:77285;C:72572;G:71816;T:97350;N:33,51,,,,77285,72572,71816,97350,33,SRX20534082,SRS17842609,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.6001,,0.12487,,0.9726,,0.56174,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75748,SRR24758312,SRX20534081,SRS17842610,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep1,GSM7432136,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep1,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432136,GSM7432136: Photoconverted brain endothelial cells rep1; Danio rerio; RNA Seq,GSM7432136 r1,GSM7432136,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049750_A01.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,250820142.0,4918042.0,GSM7432136 r1,0:51,A:64280299;C:60450772;G:60900384;T:65166579;N:22108,51,,,,64280299,60450772,60900384,65166579,22108,SRX20534081,SRS17842610,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.7733,,0.08089,,0.89792,,0.51899,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75749,SRR24758313,SRX20534081,SRS17842610,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep1,GSM7432136,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep1,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432136,GSM7432136: Photoconverted brain endothelial cells rep1; Danio rerio; RNA Seq,GSM7432136 r1,GSM7432136,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049750_A10.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,172734042.0,3386942.0,GSM7432136 r10,0:51,A:45722206;C:40210149;G:40628743;T:46158019;N:14925,51,,,,45722206,40210149,40628743,46158019,14925,SRX20534081,SRS17842610,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.81901,,0.10806,,0.88463,,0.54871,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75750,SRR24758314,SRX20534081,SRS17842610,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep1,GSM7432136,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep1,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432136,GSM7432136: Photoconverted brain endothelial cells rep1; Danio rerio; RNA Seq,GSM7432136 r1,GSM7432136,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049750_A11.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,235935537.0,4626187.0,GSM7432136 r11,0:51,A:61038358;C:56039572;G:56663106;T:62174269;N:20232,51,,,,61038358,56039572,56663106,62174269,20232,SRX20534081,SRS17842610,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.78196,,0.0733,,0.89865,,0.45906,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System 75751,SRR24758315,SRX20534081,SRS17842610,SRP440051,PRJNA977246,An organ specific angiogenic control mechanism for endothelial tailoring,GSE233662,Transcriptome Analysis,Each organ of the human body requires locally adapted blood vessels. The gain of such organotypic vessel specializations is often deemed molecularly unrelated to the process of organ vascularization. Opposing this model we reveal a molecular mechanism for brain specific angiogenesis that operates under the control of Wnt7a/b ligands well known blood brain barrier maturation signals. The control mechanism relies on Wnt7a/b dependent expression of Mmp25 in brain endothelial cells. This hitherto poorly characterized GPI anchored matrix metalloproteinase is selectively required in endothelial tip cells to enable their initial migration across the pial basement membrane lining the brain surface which distinctive molecular composition is controlled by embryonic pial fibroblasts. Mechanistically Mmp25 confers brain invasive competence by cleaving the pial basement membrane enriched Col4a5/6 within a short non collagenous region of the central helical part of the heterotrimer. Upon genetic interference with pial basement membrane composition the Wnt/ß catenin dependent organotypic control of brain angiogenesis is lost resulting in properly patterned yet blood brain barrier defective cerebrovasculatures. This work reveals an organ specific angiogenesis mechanism sheds light on tip cell mechanistic angiodiversity and thereby illustrates how organs by imposing local constraints on angiogenic tip cells can select vessels matching their distinctive physiological requirements. Overall design: The samples in this submission are single cells sorted into single wells of a 96 or 384 well plates. The cells were lysed and transcriptomic sequencing data was obtained using Smart seq2 chemistry,,pubmed:38570687,,Photoconverted brain endothelial cells rep1,GSM7432136,,source name:Brain|tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino|geo loc name:missing|collection date:missing,Photoconverted brain endothelial cells rep1,Demultiplexing with bcl2fastq Alignment against the zebrafish Danio rerio.GRCz11.95 build using Bowtie2 Removal of adapter sequencing with TrimGalore Removal of duplets using samtools Generation of a count matrix with FeatureCounts from the SubRead package Assembly: Danio.Rerio.GRCz11 Supplementary files format and content: Count files with every column a cell and every row a gene.,Brain,Morpholino injection,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank’s Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,Standard,tissue:Brain|age:30 hpf|cell type:Endothelial cells|genotype:Tgfli1a:Gal4FF;UAS:Kaede|treatment:WT and gpr124 morpholino,GSM7432136,GSM7432136: Photoconverted brain endothelial cells rep1; Danio rerio; RNA Seq,GSM7432136 r1,GSM7432136,1,Photoconversion ofTgfli1:Gal4ubs3;UAS:Kaederk8PHBC or CtAs ECs was performed using a Zeiss LSM 710 confocal microscope Carl Zeiss objective lenses: Plan Apochromat ×20/0.8 M27. Briefly anesthetised embryos were mounted laterally in 1% low melting agarose and the fluorescent Kaede protein was photoswitched by scanning the selected region of interest with a 405 nm laser five iterations of 50 s. post isolation from the agarose embryos were washed in Ca2+/Mg2+ free HBSS Hank's Balanced Salt Solution Gibco and dissociated at 28.5 °C for 30 min in TrypLE select Thermo Fischer Scientific 12563011. Dissociation was stopped by the addition of FBS and centrifugation. The cell pellet was resuspended in HBSS containing Ca2+/Mg2+and 5% FBS filtered and submitted to FACS BD Biosciences FACSAria III. Single cell cDNA libraries were prepared according to the established protocol for Smart Seq2. In brief poly adenylated mRNA was transcribed to cDNA using oligodT primer and SuperScript II reverse transcriptase ThermoFisher Scientific. Synthesis of second strand cDNA was achieved using a template switching oligo and the double stranded cDNA was then amplified using polymerase chain reaction PCR for 23 – 26 cycles. post bead purification overall cDNA quality was controlled QC by analyzing randomly selected single cell samples wells on a 2100 Bioanalyzer with a DNA High sensitivity chip Agilent Biotechnologies. When the sample plate passed the QC the cDNA was fragmented and tagged tagmented using Tn5 transposase and each single cell sample well was uniquely indexed using Illumina Nextera XT index kits set A D. Therepost the indexed single cell samples from one plate were pooled to be sequenced together on one lane of a HiSeq3000/4000 sequencer Illumina using dual indexing and 50 base read length. ScRNA Seq with Smart seq2,,RNA-Seq,TRANSCRIPTOMIC SINGLE CELL,cDNA,SINGLE,ILLUMINA,Illumina HiSeq 4000,,SRP440051,,loader:fastq load.py,GC049750_A12.180302.HiSeq4000.FCA.lane5.gcap_17_11.R1.fastq.gz,fastq,313047945.0,6138195.0,GSM7432136 r12,0:51,A:80262467;C:74589633;G:75513657;T:82654375;N:27813,51,,,,80262467,74589633,75513657,82654375,27813,SRX20534081,SRS17842610,SRA1645745,"SICOF, Department of Medicine, Huddinge, Karolinska Institute","Department of Medicine, Karolinska Institute",1,0.84159,,0.08077,,0.84971,,0.52668,,51,,B,,usable mapping rate,illumina,hiseq_era,unknown,poly_a,nextera,sc,single_cell_plate,smartseq,,Sweden,2023-05-29,Pharyngula,Embryo,Brain,Nervous System