run_metadata
1,001 rows where devstage_curation = "Undetermined" and technology = "10x"
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| Link | 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 |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 11777 | 11777 | ERR11758595 | ERX11157718 | ERS16172936 | ERP149744 | PRJEB64563 | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E-MTAB-13196 | Transcriptome Analysis | Development of the dorsal aorta is a key step in the establishment of the adult blood forming system since hematopoietic stem and progenitor cells HSPCs arise from ventral aortic endothelium in all vertebrate animals studied. Work in zebrafish has demonstrated that arterial and venous endothelial precursors arise from distinct subsets of lateral plate mesoderm. Here we profile the transcriptome of the earliest detectable endothelial cells ECs during zebrafish embryogenesis to demonstrate that tissue specific EC programs initiate much earlier than previously appreciated by the end of gastrulation. Classic studies in the chick embryo showed that paraxial mesoderm generates a subset of somite derived endothelial cells SDECs that incorporate into the dorsal aorta to replace HSPCs as they exit the aorta and enter circulation. We describe a conserved program in the zebrafish where a rare population of endothelial precursors delaminates from the dermomyotome to incorporate exclusively into the developing dorsal aorta. Whereas SDECs lack hematopoietic potential they act as a local niche to support the emergence of HSPCs from neighboring hemogenic endothelium. Thus at least three subsets of ECs contribute to the developing dorsal aorta: vascular ECs hemogenic ECs and SDECs. Taken together our findings indicate that the distinct spatial origins of endothelial precursors dictate different cellular potentials within the developing dorsal aorta. | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | Protocols: Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | drl tb | SAMEA114192243 | Institute of Molecular Genetics of the Czech Academy of Sciences | ENA FIRST PUBLIC:2023 08 01T00:17:16Z|ENA LAST UPDATE:2023 08 01T00:17:16Z|External Id:SAMEA114192243|INSDC center alias:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC center name:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC first public:2023 08 01T00:17:16Z|INSDC last update:2023 08 01T00:17:16Z|INSDC status:public|Submitter Id:E MTAB 13196:drl tb|age:10|broker name:ArrayExpress|cell type:endothelial cell|collection date:not collected|common name:zebrafish|developmental stage:tail bud prominent stage|genotype:drl:H2B Dendra2|geographic location country and/or sea:not collected|immunophenotype:drl:H2B Dendra2+|isolate:not applicable|organism part:embryo|sample name:E MTAB 13196:drl tb|scientific name:Danio rerio|strain:drl:H2B Dendra2 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E MTAB 13196:drl tb p | drl tb p | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | Oligo-dT | PAIRED | ILLUMINA | Illumina HiSeq 2500 | ERP149744 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | drl_h2b-dendra_tb_S36_L001_R2_001.fastq.gz drl_h2b-dendra_tb_S36_L001_R1_001.fastq.gz | fastq fastq | 211593816.0 | 1679316.0 | E MTAB 13196:drl h2b dendra tb S36 L001 | 0:28 1:98 | A:62689430;C:45809168;G:45709128;T:57272091;N:113999 | 28 | 98 | 62689430 | 45809168 | 45709128 | 57272091 | 113999 | ERX11157718 | ERS16172936 | ERA25628220 | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | 2 | 0.00192 | 0.91331 | 0.00072 | 0.14588 | 0.99675 | 0.89292 | 0.51304 | 0.63144 | 28 | 98 | T | B | sc-like readlen | illumina | hiseq_era | unknown | poly_a | unknown | sc | single_cell_droplet | 10x | Czech Republic | 2023-08-01 | Undetermined | Embryo | Embryo Imprecise | All anatomical structures | ||||||||||||||
| 11778 | 11778 | ERR11758629 | ERX11157718 | ERS16172936 | ERP149744 | PRJEB64563 | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E-MTAB-13196 | Transcriptome Analysis | Development of the dorsal aorta is a key step in the establishment of the adult blood forming system since hematopoietic stem and progenitor cells HSPCs arise from ventral aortic endothelium in all vertebrate animals studied. Work in zebrafish has demonstrated that arterial and venous endothelial precursors arise from distinct subsets of lateral plate mesoderm. Here we profile the transcriptome of the earliest detectable endothelial cells ECs during zebrafish embryogenesis to demonstrate that tissue specific EC programs initiate much earlier than previously appreciated by the end of gastrulation. Classic studies in the chick embryo showed that paraxial mesoderm generates a subset of somite derived endothelial cells SDECs that incorporate into the dorsal aorta to replace HSPCs as they exit the aorta and enter circulation. We describe a conserved program in the zebrafish where a rare population of endothelial precursors delaminates from the dermomyotome to incorporate exclusively into the developing dorsal aorta. Whereas SDECs lack hematopoietic potential they act as a local niche to support the emergence of HSPCs from neighboring hemogenic endothelium. Thus at least three subsets of ECs contribute to the developing dorsal aorta: vascular ECs hemogenic ECs and SDECs. Taken together our findings indicate that the distinct spatial origins of endothelial precursors dictate different cellular potentials within the developing dorsal aorta. | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | Protocols: Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | drl tb | SAMEA114192243 | Institute of Molecular Genetics of the Czech Academy of Sciences | ENA FIRST PUBLIC:2023 08 01T00:17:16Z|ENA LAST UPDATE:2023 08 01T00:17:16Z|External Id:SAMEA114192243|INSDC center alias:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC center name:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC first public:2023 08 01T00:17:16Z|INSDC last update:2023 08 01T00:17:16Z|INSDC status:public|Submitter Id:E MTAB 13196:drl tb|age:10|broker name:ArrayExpress|cell type:endothelial cell|collection date:not collected|common name:zebrafish|developmental stage:tail bud prominent stage|genotype:drl:H2B Dendra2|geographic location country and/or sea:not collected|immunophenotype:drl:H2B Dendra2+|isolate:not applicable|organism part:embryo|sample name:E MTAB 13196:drl tb|scientific name:Danio rerio|strain:drl:H2B Dendra2 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E MTAB 13196:drl tb p | drl tb p | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | Oligo-dT | PAIRED | ILLUMINA | Illumina HiSeq 2500 | ERP149744 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | drl_h2b-dendra_tb_S37_L001_R2_001.fastq.gz drl_h2b-dendra_tb_S37_L001_R1_001.fastq.gz | fastq fastq | 980164836.0 | 7779086.0 | E MTAB 13196:drl h2b dendra tb S37 L001 | 0:28 1:98 | A:281375011;C:216067640;G:222839674;T:259781180;N:101331 | 28 | 98 | 281375011 | 216067640 | 222839674 | 259781180 | 101331 | ERX11157718 | ERS16172936 | ERA25628220 | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | 2 | 0.00817 | 0.89718 | 0.00237 | 0.12153 | 0.98912 | 0.82643 | 0.46403 | 0.64954 | 28 | 98 | T | B | sc-like readlen | illumina | hiseq_era | unknown | poly_a | unknown | sc | single_cell_droplet | 10x | Czech Republic | 2023-08-01 | Undetermined | Embryo | Embryo Imprecise | All anatomical structures | ||||||||||||||
| 11779 | 11779 | ERR11758618 | ERX11157718 | ERS16172936 | ERP149744 | PRJEB64563 | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E-MTAB-13196 | Transcriptome Analysis | Development of the dorsal aorta is a key step in the establishment of the adult blood forming system since hematopoietic stem and progenitor cells HSPCs arise from ventral aortic endothelium in all vertebrate animals studied. Work in zebrafish has demonstrated that arterial and venous endothelial precursors arise from distinct subsets of lateral plate mesoderm. Here we profile the transcriptome of the earliest detectable endothelial cells ECs during zebrafish embryogenesis to demonstrate that tissue specific EC programs initiate much earlier than previously appreciated by the end of gastrulation. Classic studies in the chick embryo showed that paraxial mesoderm generates a subset of somite derived endothelial cells SDECs that incorporate into the dorsal aorta to replace HSPCs as they exit the aorta and enter circulation. We describe a conserved program in the zebrafish where a rare population of endothelial precursors delaminates from the dermomyotome to incorporate exclusively into the developing dorsal aorta. Whereas SDECs lack hematopoietic potential they act as a local niche to support the emergence of HSPCs from neighboring hemogenic endothelium. Thus at least three subsets of ECs contribute to the developing dorsal aorta: vascular ECs hemogenic ECs and SDECs. Taken together our findings indicate that the distinct spatial origins of endothelial precursors dictate different cellular potentials within the developing dorsal aorta. | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | Protocols: Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | drl tb | SAMEA114192243 | Institute of Molecular Genetics of the Czech Academy of Sciences | ENA FIRST PUBLIC:2023 08 01T00:17:16Z|ENA LAST UPDATE:2023 08 01T00:17:16Z|External Id:SAMEA114192243|INSDC center alias:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC center name:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC first public:2023 08 01T00:17:16Z|INSDC last update:2023 08 01T00:17:16Z|INSDC status:public|Submitter Id:E MTAB 13196:drl tb|age:10|broker name:ArrayExpress|cell type:endothelial cell|collection date:not collected|common name:zebrafish|developmental stage:tail bud prominent stage|genotype:drl:H2B Dendra2|geographic location country and/or sea:not collected|immunophenotype:drl:H2B Dendra2+|isolate:not applicable|organism part:embryo|sample name:E MTAB 13196:drl tb|scientific name:Danio rerio|strain:drl:H2B Dendra2 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E MTAB 13196:drl tb p | drl tb p | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | Oligo-dT | PAIRED | ILLUMINA | Illumina HiSeq 2500 | ERP149744 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | drl_h2b-dendra_tb_S39_L001_R2_001.fastq.gz drl_h2b-dendra_tb_S39_L001_R1_001.fastq.gz | fastq fastq | 12118302.0 | 96177.0 | E MTAB 13196:drl h2b dendra tb S39 L001 | 0:28 1:98 | A:3543790;C:2662872;G:2728134;T:3182383;N:1123 | 28 | 98 | 3543790 | 2662872 | 2728134 | 3182383 | 1123 | ERX11157718 | ERS16172936 | ERA25628220 | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | 2 | 0.00786 | 0.88963 | 0.0025 | 0.12543 | 0.99403 | 0.88635 | 0.50098 | 0.64694 | 28 | 98 | T | B | sc-like readlen | illumina | hiseq_era | unknown | poly_a | unknown | sc | single_cell_droplet | 10x | Czech Republic | 2023-08-01 | Undetermined | Embryo | Embryo Imprecise | All anatomical structures | ||||||||||||||
| 11780 | 11780 | ERR11758643 | ERX11157718 | ERS16172936 | ERP149744 | PRJEB64563 | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E-MTAB-13196 | Transcriptome Analysis | Development of the dorsal aorta is a key step in the establishment of the adult blood forming system since hematopoietic stem and progenitor cells HSPCs arise from ventral aortic endothelium in all vertebrate animals studied. Work in zebrafish has demonstrated that arterial and venous endothelial precursors arise from distinct subsets of lateral plate mesoderm. Here we profile the transcriptome of the earliest detectable endothelial cells ECs during zebrafish embryogenesis to demonstrate that tissue specific EC programs initiate much earlier than previously appreciated by the end of gastrulation. Classic studies in the chick embryo showed that paraxial mesoderm generates a subset of somite derived endothelial cells SDECs that incorporate into the dorsal aorta to replace HSPCs as they exit the aorta and enter circulation. We describe a conserved program in the zebrafish where a rare population of endothelial precursors delaminates from the dermomyotome to incorporate exclusively into the developing dorsal aorta. Whereas SDECs lack hematopoietic potential they act as a local niche to support the emergence of HSPCs from neighboring hemogenic endothelium. Thus at least three subsets of ECs contribute to the developing dorsal aorta: vascular ECs hemogenic ECs and SDECs. Taken together our findings indicate that the distinct spatial origins of endothelial precursors dictate different cellular potentials within the developing dorsal aorta. | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | Protocols: Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | drl tb | SAMEA114192243 | Institute of Molecular Genetics of the Czech Academy of Sciences | ENA FIRST PUBLIC:2023 08 01T00:17:16Z|ENA LAST UPDATE:2023 08 01T00:17:16Z|External Id:SAMEA114192243|INSDC center alias:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC center name:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC first public:2023 08 01T00:17:16Z|INSDC last update:2023 08 01T00:17:16Z|INSDC status:public|Submitter Id:E MTAB 13196:drl tb|age:10|broker name:ArrayExpress|cell type:endothelial cell|collection date:not collected|common name:zebrafish|developmental stage:tail bud prominent stage|genotype:drl:H2B Dendra2|geographic location country and/or sea:not collected|immunophenotype:drl:H2B Dendra2+|isolate:not applicable|organism part:embryo|sample name:E MTAB 13196:drl tb|scientific name:Danio rerio|strain:drl:H2B Dendra2 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E MTAB 13196:drl tb p | drl tb p | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | Oligo-dT | PAIRED | ILLUMINA | Illumina HiSeq 2500 | ERP149744 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | drl_h2b-dendra_tb_S5_L005_R2_001.fastq.gz drl_h2b-dendra_tb_S5_L005_R1_001.fastq.gz | fastq fastq | 2061022698.0 | 16357323.0 | E MTAB 13196:drl h2b dendra tb S5 L005 | 0:28 1:98 | A:587730953;C:457631700;G:470283474;T:545196979;N:179592 | 28 | 98 | 587730953 | 457631700 | 470283474 | 545196979 | 179592 | ERX11157718 | ERS16172936 | ERA25628220 | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | 2 | 0.01192 | 0.91661 | 0.00336 | 0.12397 | 0.98518 | 0.82568 | 0.45692 | 0.65555 | 28 | 98 | T | B | sc-like readlen | illumina | hiseq_era | unknown | poly_a | unknown | sc | single_cell_droplet | 10x | Czech Republic | 2023-08-01 | Undetermined | Embryo | Embryo Imprecise | All anatomical structures | ||||||||||||||
| 11781 | 11781 | ERR11758610 | ERX11157718 | ERS16172936 | ERP149744 | PRJEB64563 | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E-MTAB-13196 | Transcriptome Analysis | Development of the dorsal aorta is a key step in the establishment of the adult blood forming system since hematopoietic stem and progenitor cells HSPCs arise from ventral aortic endothelium in all vertebrate animals studied. Work in zebrafish has demonstrated that arterial and venous endothelial precursors arise from distinct subsets of lateral plate mesoderm. Here we profile the transcriptome of the earliest detectable endothelial cells ECs during zebrafish embryogenesis to demonstrate that tissue specific EC programs initiate much earlier than previously appreciated by the end of gastrulation. Classic studies in the chick embryo showed that paraxial mesoderm generates a subset of somite derived endothelial cells SDECs that incorporate into the dorsal aorta to replace HSPCs as they exit the aorta and enter circulation. We describe a conserved program in the zebrafish where a rare population of endothelial precursors delaminates from the dermomyotome to incorporate exclusively into the developing dorsal aorta. Whereas SDECs lack hematopoietic potential they act as a local niche to support the emergence of HSPCs from neighboring hemogenic endothelium. Thus at least three subsets of ECs contribute to the developing dorsal aorta: vascular ECs hemogenic ECs and SDECs. Taken together our findings indicate that the distinct spatial origins of endothelial precursors dictate different cellular potentials within the developing dorsal aorta. | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | Protocols: Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | drl tb | SAMEA114192243 | Institute of Molecular Genetics of the Czech Academy of Sciences | ENA FIRST PUBLIC:2023 08 01T00:17:16Z|ENA LAST UPDATE:2023 08 01T00:17:16Z|External Id:SAMEA114192243|INSDC center alias:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC center name:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC first public:2023 08 01T00:17:16Z|INSDC last update:2023 08 01T00:17:16Z|INSDC status:public|Submitter Id:E MTAB 13196:drl tb|age:10|broker name:ArrayExpress|cell type:endothelial cell|collection date:not collected|common name:zebrafish|developmental stage:tail bud prominent stage|genotype:drl:H2B Dendra2|geographic location country and/or sea:not collected|immunophenotype:drl:H2B Dendra2+|isolate:not applicable|organism part:embryo|sample name:E MTAB 13196:drl tb|scientific name:Danio rerio|strain:drl:H2B Dendra2 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E MTAB 13196:drl tb p | drl tb p | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | Oligo-dT | PAIRED | ILLUMINA | Illumina HiSeq 2500 | ERP149744 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | drl_h2b-dendra_tb_S38_L001_R2_001.fastq.gz drl_h2b-dendra_tb_S38_L001_R1_001.fastq.gz | fastq fastq | 924603120.0 | 7338120.0 | E MTAB 13196:drl h2b dendra tb S38 L001 | 0:28 1:98 | A:265987815;C:203824483;G:210024160;T:244670646;N:96016 | 28 | 98 | 265987815 | 203824483 | 210024160 | 244670646 | 96016 | ERX11157718 | ERS16172936 | ERA25628220 | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | 2 | 0.00773 | 0.89881 | 0.00242 | 0.12145 | 0.98944 | 0.82948 | 0.50529 | 0.66568 | 28 | 98 | T | B | sc-like readlen | illumina | hiseq_era | unknown | poly_a | unknown | sc | single_cell_droplet | 10x | Czech Republic | 2023-08-01 | Undetermined | Embryo | Embryo Imprecise | All anatomical structures | ||||||||||||||
| 11782 | 11782 | ERR11758641 | ERX11157718 | ERS16172936 | ERP149744 | PRJEB64563 | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E-MTAB-13196 | Transcriptome Analysis | Development of the dorsal aorta is a key step in the establishment of the adult blood forming system since hematopoietic stem and progenitor cells HSPCs arise from ventral aortic endothelium in all vertebrate animals studied. Work in zebrafish has demonstrated that arterial and venous endothelial precursors arise from distinct subsets of lateral plate mesoderm. Here we profile the transcriptome of the earliest detectable endothelial cells ECs during zebrafish embryogenesis to demonstrate that tissue specific EC programs initiate much earlier than previously appreciated by the end of gastrulation. Classic studies in the chick embryo showed that paraxial mesoderm generates a subset of somite derived endothelial cells SDECs that incorporate into the dorsal aorta to replace HSPCs as they exit the aorta and enter circulation. We describe a conserved program in the zebrafish where a rare population of endothelial precursors delaminates from the dermomyotome to incorporate exclusively into the developing dorsal aorta. Whereas SDECs lack hematopoietic potential they act as a local niche to support the emergence of HSPCs from neighboring hemogenic endothelium. Thus at least three subsets of ECs contribute to the developing dorsal aorta: vascular ECs hemogenic ECs and SDECs. Taken together our findings indicate that the distinct spatial origins of endothelial precursors dictate different cellular potentials within the developing dorsal aorta. | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | Protocols: Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | drl tb | SAMEA114192243 | Institute of Molecular Genetics of the Czech Academy of Sciences | ENA FIRST PUBLIC:2023 08 01T00:17:16Z|ENA LAST UPDATE:2023 08 01T00:17:16Z|External Id:SAMEA114192243|INSDC center alias:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC center name:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC first public:2023 08 01T00:17:16Z|INSDC last update:2023 08 01T00:17:16Z|INSDC status:public|Submitter Id:E MTAB 13196:drl tb|age:10|broker name:ArrayExpress|cell type:endothelial cell|collection date:not collected|common name:zebrafish|developmental stage:tail bud prominent stage|genotype:drl:H2B Dendra2|geographic location country and/or sea:not collected|immunophenotype:drl:H2B Dendra2+|isolate:not applicable|organism part:embryo|sample name:E MTAB 13196:drl tb|scientific name:Danio rerio|strain:drl:H2B Dendra2 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E MTAB 13196:drl tb p | drl tb p | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | Oligo-dT | PAIRED | ILLUMINA | Illumina HiSeq 2500 | ERP149744 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | drl_h2b-dendra_tb_S6_L005_R2_001.fastq.gz drl_h2b-dendra_tb_S6_L005_R1_001.fastq.gz | fastq fastq | 1954661562.0 | 15513187.0 | E MTAB 13196:drl h2b dendra tb S6 L005 | 0:28 1:98 | A:558769608;C:433724557;G:445550100;T:516443984;N:173313 | 28 | 98 | 558769608 | 433724557 | 445550100 | 516443984 | 173313 | ERX11157718 | ERS16172936 | ERA25628220 | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | 2 | 0.0125 | 0.91697 | 0.00373 | 0.12359 | 0.98447 | 0.82558 | 0.4881 | 0.6456 | 28 | 98 | T | B | sc-like readlen | illumina | hiseq_era | unknown | poly_a | unknown | sc | single_cell_droplet | 10x | Czech Republic | 2023-08-01 | Undetermined | Embryo | Embryo Imprecise | All anatomical structures | ||||||||||||||
| 11783 | 11783 | ERR11758591 | ERX11157718 | ERS16172936 | ERP149744 | PRJEB64563 | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E-MTAB-13196 | Transcriptome Analysis | Development of the dorsal aorta is a key step in the establishment of the adult blood forming system since hematopoietic stem and progenitor cells HSPCs arise from ventral aortic endothelium in all vertebrate animals studied. Work in zebrafish has demonstrated that arterial and venous endothelial precursors arise from distinct subsets of lateral plate mesoderm. Here we profile the transcriptome of the earliest detectable endothelial cells ECs during zebrafish embryogenesis to demonstrate that tissue specific EC programs initiate much earlier than previously appreciated by the end of gastrulation. Classic studies in the chick embryo showed that paraxial mesoderm generates a subset of somite derived endothelial cells SDECs that incorporate into the dorsal aorta to replace HSPCs as they exit the aorta and enter circulation. We describe a conserved program in the zebrafish where a rare population of endothelial precursors delaminates from the dermomyotome to incorporate exclusively into the developing dorsal aorta. Whereas SDECs lack hematopoietic potential they act as a local niche to support the emergence of HSPCs from neighboring hemogenic endothelium. Thus at least three subsets of ECs contribute to the developing dorsal aorta: vascular ECs hemogenic ECs and SDECs. Taken together our findings indicate that the distinct spatial origins of endothelial precursors dictate different cellular potentials within the developing dorsal aorta. | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | Protocols: Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | drl tb | SAMEA114192243 | Institute of Molecular Genetics of the Czech Academy of Sciences | ENA FIRST PUBLIC:2023 08 01T00:17:16Z|ENA LAST UPDATE:2023 08 01T00:17:16Z|External Id:SAMEA114192243|INSDC center alias:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC center name:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC first public:2023 08 01T00:17:16Z|INSDC last update:2023 08 01T00:17:16Z|INSDC status:public|Submitter Id:E MTAB 13196:drl tb|age:10|broker name:ArrayExpress|cell type:endothelial cell|collection date:not collected|common name:zebrafish|developmental stage:tail bud prominent stage|genotype:drl:H2B Dendra2|geographic location country and/or sea:not collected|immunophenotype:drl:H2B Dendra2+|isolate:not applicable|organism part:embryo|sample name:E MTAB 13196:drl tb|scientific name:Danio rerio|strain:drl:H2B Dendra2 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E MTAB 13196:drl tb p | drl tb p | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | Oligo-dT | PAIRED | ILLUMINA | Illumina HiSeq 2500 | ERP149744 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | drl_h2b-dendra_tb_S35_L001_R2_001.fastq.gz drl_h2b-dendra_tb_S35_L001_R1_001.fastq.gz | fastq fastq | 4717188.0 | 37438.0 | E MTAB 13196:drl h2b dendra tb S35 L001 | 0:28 1:98 | A:1419430;C:1014265;G:1016661;T:1264642;N:2190 | 28 | 98 | 1419430 | 1014265 | 1016661 | 1264642 | 2190 | ERX11157718 | ERS16172936 | ERA25628220 | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | 2 | 0.0012 | 0.65482 | 0.00054 | 0.10783 | 0.99953 | 0.97157 | 0.625 | 0.63689 | 28 | 98 | T | B | sc-like readlen | illumina | hiseq_era | unknown | poly_a | unknown | sc | single_cell_droplet | 10x | Czech Republic | 2023-08-01 | Undetermined | Embryo | Embryo Imprecise | All anatomical structures | ||||||||||||||
| 11784 | 11784 | ERR11758606 | ERX11157718 | ERS16172936 | ERP149744 | PRJEB64563 | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E-MTAB-13196 | Transcriptome Analysis | Development of the dorsal aorta is a key step in the establishment of the adult blood forming system since hematopoietic stem and progenitor cells HSPCs arise from ventral aortic endothelium in all vertebrate animals studied. Work in zebrafish has demonstrated that arterial and venous endothelial precursors arise from distinct subsets of lateral plate mesoderm. Here we profile the transcriptome of the earliest detectable endothelial cells ECs during zebrafish embryogenesis to demonstrate that tissue specific EC programs initiate much earlier than previously appreciated by the end of gastrulation. Classic studies in the chick embryo showed that paraxial mesoderm generates a subset of somite derived endothelial cells SDECs that incorporate into the dorsal aorta to replace HSPCs as they exit the aorta and enter circulation. We describe a conserved program in the zebrafish where a rare population of endothelial precursors delaminates from the dermomyotome to incorporate exclusively into the developing dorsal aorta. Whereas SDECs lack hematopoietic potential they act as a local niche to support the emergence of HSPCs from neighboring hemogenic endothelium. Thus at least three subsets of ECs contribute to the developing dorsal aorta: vascular ECs hemogenic ECs and SDECs. Taken together our findings indicate that the distinct spatial origins of endothelial precursors dictate different cellular potentials within the developing dorsal aorta. | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | Protocols: Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | drl tb | SAMEA114192243 | Institute of Molecular Genetics of the Czech Academy of Sciences | ENA FIRST PUBLIC:2023 08 01T00:17:16Z|ENA LAST UPDATE:2023 08 01T00:17:16Z|External Id:SAMEA114192243|INSDC center alias:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC center name:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC first public:2023 08 01T00:17:16Z|INSDC last update:2023 08 01T00:17:16Z|INSDC status:public|Submitter Id:E MTAB 13196:drl tb|age:10|broker name:ArrayExpress|cell type:endothelial cell|collection date:not collected|common name:zebrafish|developmental stage:tail bud prominent stage|genotype:drl:H2B Dendra2|geographic location country and/or sea:not collected|immunophenotype:drl:H2B Dendra2+|isolate:not applicable|organism part:embryo|sample name:E MTAB 13196:drl tb|scientific name:Danio rerio|strain:drl:H2B Dendra2 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E MTAB 13196:drl tb p | drl tb p | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | Oligo-dT | PAIRED | ILLUMINA | Illumina HiSeq 2500 | ERP149744 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | drl_h2b-dendra_tb_S34_L001_R1_001.fastq.gz drl_h2b-dendra_tb_S34_L001_R2_001.fastq.gz | fastq fastq | 303645636.0 | 2409886.0 | E MTAB 13196:drl h2b dendra tb S34 L001 | 0:28 1:98 | A:90748559;C:65369482;G:65969002;T:81405258;N:153335 | 28 | 98 | 90748559 | 65369482 | 65969002 | 81405258 | 153335 | ERX11157718 | ERS16172936 | ERA25628220 | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | 2 | 0.00228 | 0.91262 | 0.00099 | 0.14666 | 0.99638 | 0.89006 | 0.47177 | 0.63109 | 28 | 98 | T | B | sc-like readlen | illumina | hiseq_era | unknown | poly_a | unknown | sc | single_cell_droplet | 10x | Czech Republic | 2023-08-01 | Undetermined | Embryo | Embryo Imprecise | All anatomical structures | ||||||||||||||
| 11785 | 11785 | ERR11758640 | ERX11157718 | ERS16172936 | ERP149744 | PRJEB64563 | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E-MTAB-13196 | Transcriptome Analysis | Development of the dorsal aorta is a key step in the establishment of the adult blood forming system since hematopoietic stem and progenitor cells HSPCs arise from ventral aortic endothelium in all vertebrate animals studied. Work in zebrafish has demonstrated that arterial and venous endothelial precursors arise from distinct subsets of lateral plate mesoderm. Here we profile the transcriptome of the earliest detectable endothelial cells ECs during zebrafish embryogenesis to demonstrate that tissue specific EC programs initiate much earlier than previously appreciated by the end of gastrulation. Classic studies in the chick embryo showed that paraxial mesoderm generates a subset of somite derived endothelial cells SDECs that incorporate into the dorsal aorta to replace HSPCs as they exit the aorta and enter circulation. We describe a conserved program in the zebrafish where a rare population of endothelial precursors delaminates from the dermomyotome to incorporate exclusively into the developing dorsal aorta. Whereas SDECs lack hematopoietic potential they act as a local niche to support the emergence of HSPCs from neighboring hemogenic endothelium. Thus at least three subsets of ECs contribute to the developing dorsal aorta: vascular ECs hemogenic ECs and SDECs. Taken together our findings indicate that the distinct spatial origins of endothelial precursors dictate different cellular potentials within the developing dorsal aorta. | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | Protocols: Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | drl tb | SAMEA114192243 | Institute of Molecular Genetics of the Czech Academy of Sciences | ENA FIRST PUBLIC:2023 08 01T00:17:16Z|ENA LAST UPDATE:2023 08 01T00:17:16Z|External Id:SAMEA114192243|INSDC center alias:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC center name:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC first public:2023 08 01T00:17:16Z|INSDC last update:2023 08 01T00:17:16Z|INSDC status:public|Submitter Id:E MTAB 13196:drl tb|age:10|broker name:ArrayExpress|cell type:endothelial cell|collection date:not collected|common name:zebrafish|developmental stage:tail bud prominent stage|genotype:drl:H2B Dendra2|geographic location country and/or sea:not collected|immunophenotype:drl:H2B Dendra2+|isolate:not applicable|organism part:embryo|sample name:E MTAB 13196:drl tb|scientific name:Danio rerio|strain:drl:H2B Dendra2 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E MTAB 13196:drl tb p | drl tb p | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | Oligo-dT | PAIRED | ILLUMINA | Illumina HiSeq 2500 | ERP149744 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | drl_h2b-dendra_tb_S40_L001_R2_001.fastq.gz drl_h2b-dendra_tb_S40_L001_R1_001.fastq.gz | fastq fastq | 661361022.0 | 5248897.0 | E MTAB 13196:drl h2b dendra tb S40 L001 | 0:28 1:98 | A:190087869;C:145985661;G:150124621;T:175095762;N:67109 | 28 | 98 | 190087869 | 145985661 | 150124621 | 175095762 | 67109 | ERX11157718 | ERS16172936 | ERA25628220 | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | 2 | 0.00785 | 0.89784 | 0.00251 | 0.12159 | 0.98948 | 0.83055 | 0.46095 | 0.64951 | 28 | 98 | T | B | sc-like readlen | illumina | hiseq_era | unknown | poly_a | unknown | sc | single_cell_droplet | 10x | Czech Republic | 2023-08-01 | Undetermined | Embryo | Embryo Imprecise | All anatomical structures | ||||||||||||||
| 11786 | 11786 | ERR11758605 | ERX11157718 | ERS16172936 | ERP149744 | PRJEB64563 | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E-MTAB-13196 | Transcriptome Analysis | Development of the dorsal aorta is a key step in the establishment of the adult blood forming system since hematopoietic stem and progenitor cells HSPCs arise from ventral aortic endothelium in all vertebrate animals studied. Work in zebrafish has demonstrated that arterial and venous endothelial precursors arise from distinct subsets of lateral plate mesoderm. Here we profile the transcriptome of the earliest detectable endothelial cells ECs during zebrafish embryogenesis to demonstrate that tissue specific EC programs initiate much earlier than previously appreciated by the end of gastrulation. Classic studies in the chick embryo showed that paraxial mesoderm generates a subset of somite derived endothelial cells SDECs that incorporate into the dorsal aorta to replace HSPCs as they exit the aorta and enter circulation. We describe a conserved program in the zebrafish where a rare population of endothelial precursors delaminates from the dermomyotome to incorporate exclusively into the developing dorsal aorta. Whereas SDECs lack hematopoietic potential they act as a local niche to support the emergence of HSPCs from neighboring hemogenic endothelium. Thus at least three subsets of ECs contribute to the developing dorsal aorta: vascular ECs hemogenic ECs and SDECs. Taken together our findings indicate that the distinct spatial origins of endothelial precursors dictate different cellular potentials within the developing dorsal aorta. | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | Protocols: Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | drl tb | SAMEA114192243 | Institute of Molecular Genetics of the Czech Academy of Sciences | ENA FIRST PUBLIC:2023 08 01T00:17:16Z|ENA LAST UPDATE:2023 08 01T00:17:16Z|External Id:SAMEA114192243|INSDC center alias:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC center name:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC first public:2023 08 01T00:17:16Z|INSDC last update:2023 08 01T00:17:16Z|INSDC status:public|Submitter Id:E MTAB 13196:drl tb|age:10|broker name:ArrayExpress|cell type:endothelial cell|collection date:not collected|common name:zebrafish|developmental stage:tail bud prominent stage|genotype:drl:H2B Dendra2|geographic location country and/or sea:not collected|immunophenotype:drl:H2B Dendra2+|isolate:not applicable|organism part:embryo|sample name:E MTAB 13196:drl tb|scientific name:Danio rerio|strain:drl:H2B Dendra2 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E MTAB 13196:drl tb p | drl tb p | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | Oligo-dT | PAIRED | ILLUMINA | Illumina HiSeq 2500 | ERP149744 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | drl_h2b-dendra_tb_S7_L005_R2_001.fastq.gz drl_h2b-dendra_tb_S7_L005_R1_001.fastq.gz | fastq fastq | 25157790.0 | 199665.0 | E MTAB 13196:drl h2b dendra tb S7 L005 | 0:28 1:98 | A:7330053;C:5600451;G:5635840;T:6589498;N:1948 | 28 | 98 | 7330053 | 5600451 | 5635840 | 6589498 | 1948 | ERX11157718 | ERS16172936 | ERA25628220 | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | 2 | 0.01201 | 0.91385 | 0.00341 | 0.12826 | 0.98595 | 0.85161 | 0.49226 | 0.64953 | 28 | 98 | T | B | sc-like readlen | illumina | hiseq_era | unknown | poly_a | unknown | sc | single_cell_droplet | 10x | Czech Republic | 2023-08-01 | Undetermined | Embryo | Embryo Imprecise | All anatomical structures | ||||||||||||||
| 11787 | 11787 | ERR11758600 | ERX11157718 | ERS16172936 | ERP149744 | PRJEB64563 | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E-MTAB-13196 | Transcriptome Analysis | Development of the dorsal aorta is a key step in the establishment of the adult blood forming system since hematopoietic stem and progenitor cells HSPCs arise from ventral aortic endothelium in all vertebrate animals studied. Work in zebrafish has demonstrated that arterial and venous endothelial precursors arise from distinct subsets of lateral plate mesoderm. Here we profile the transcriptome of the earliest detectable endothelial cells ECs during zebrafish embryogenesis to demonstrate that tissue specific EC programs initiate much earlier than previously appreciated by the end of gastrulation. Classic studies in the chick embryo showed that paraxial mesoderm generates a subset of somite derived endothelial cells SDECs that incorporate into the dorsal aorta to replace HSPCs as they exit the aorta and enter circulation. We describe a conserved program in the zebrafish where a rare population of endothelial precursors delaminates from the dermomyotome to incorporate exclusively into the developing dorsal aorta. Whereas SDECs lack hematopoietic potential they act as a local niche to support the emergence of HSPCs from neighboring hemogenic endothelium. Thus at least three subsets of ECs contribute to the developing dorsal aorta: vascular ECs hemogenic ECs and SDECs. Taken together our findings indicate that the distinct spatial origins of endothelial precursors dictate different cellular potentials within the developing dorsal aorta. | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | Protocols: Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | drl tb | SAMEA114192243 | Institute of Molecular Genetics of the Czech Academy of Sciences | ENA FIRST PUBLIC:2023 08 01T00:17:16Z|ENA LAST UPDATE:2023 08 01T00:17:16Z|External Id:SAMEA114192243|INSDC center alias:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC center name:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC first public:2023 08 01T00:17:16Z|INSDC last update:2023 08 01T00:17:16Z|INSDC status:public|Submitter Id:E MTAB 13196:drl tb|age:10|broker name:ArrayExpress|cell type:endothelial cell|collection date:not collected|common name:zebrafish|developmental stage:tail bud prominent stage|genotype:drl:H2B Dendra2|geographic location country and/or sea:not collected|immunophenotype:drl:H2B Dendra2+|isolate:not applicable|organism part:embryo|sample name:E MTAB 13196:drl tb|scientific name:Danio rerio|strain:drl:H2B Dendra2 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E MTAB 13196:drl tb p | drl tb p | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | Oligo-dT | PAIRED | ILLUMINA | Illumina HiSeq 2500 | ERP149744 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | drl_h2b-dendra_tb_S8_L005_R2_001.fastq.gz drl_h2b-dendra_tb_S8_L005_R1_001.fastq.gz | fastq fastq | 1422735804.0 | 11291554.0 | E MTAB 13196:drl h2b dendra tb S8 L005 | 0:28 1:98 | A:405788442;C:317299433;G:322206476;T:377317416;N:124037 | 28 | 98 | 405788442 | 317299433 | 322206476 | 377317416 | 124037 | ERX11157718 | ERS16172936 | ERA25628220 | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | 2 | 0.01226 | 0.91547 | 0.0037 | 0.12565 | 0.9849 | 0.83662 | 0.48279 | 0.64531 | 28 | 98 | T | B | sc-like readlen | illumina | hiseq_era | unknown | poly_a | unknown | sc | single_cell_droplet | 10x | Czech Republic | 2023-08-01 | Undetermined | Embryo | Embryo Imprecise | All anatomical structures | ||||||||||||||
| 11788 | 11788 | ERR11758616 | ERX11157718 | ERS16172936 | ERP149744 | PRJEB64563 | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E-MTAB-13196 | Transcriptome Analysis | Development of the dorsal aorta is a key step in the establishment of the adult blood forming system since hematopoietic stem and progenitor cells HSPCs arise from ventral aortic endothelium in all vertebrate animals studied. Work in zebrafish has demonstrated that arterial and venous endothelial precursors arise from distinct subsets of lateral plate mesoderm. Here we profile the transcriptome of the earliest detectable endothelial cells ECs during zebrafish embryogenesis to demonstrate that tissue specific EC programs initiate much earlier than previously appreciated by the end of gastrulation. Classic studies in the chick embryo showed that paraxial mesoderm generates a subset of somite derived endothelial cells SDECs that incorporate into the dorsal aorta to replace HSPCs as they exit the aorta and enter circulation. We describe a conserved program in the zebrafish where a rare population of endothelial precursors delaminates from the dermomyotome to incorporate exclusively into the developing dorsal aorta. Whereas SDECs lack hematopoietic potential they act as a local niche to support the emergence of HSPCs from neighboring hemogenic endothelium. Thus at least three subsets of ECs contribute to the developing dorsal aorta: vascular ECs hemogenic ECs and SDECs. Taken together our findings indicate that the distinct spatial origins of endothelial precursors dictate different cellular potentials within the developing dorsal aorta. | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | Protocols: Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3ʹ GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | drl tb | SAMEA114192243 | Institute of Molecular Genetics of the Czech Academy of Sciences | ENA FIRST PUBLIC:2023 08 01T00:17:16Z|ENA LAST UPDATE:2023 08 01T00:17:16Z|External Id:SAMEA114192243|INSDC center alias:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC center name:Institute of Molecular Genetics of the Czech Academy of Sciences|INSDC first public:2023 08 01T00:17:16Z|INSDC last update:2023 08 01T00:17:16Z|INSDC status:public|Submitter Id:E MTAB 13196:drl tb|age:10|broker name:ArrayExpress|cell type:endothelial cell|collection date:not collected|common name:zebrafish|developmental stage:tail bud prominent stage|genotype:drl:H2B Dendra2|geographic location country and/or sea:not collected|immunophenotype:drl:H2B Dendra2+|isolate:not applicable|organism part:embryo|sample name:E MTAB 13196:drl tb|scientific name:Danio rerio|strain:drl:H2B Dendra2 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | E MTAB 13196:drl tb p | drl tb p | Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | Dechorionated embryos were collected at defined stages dissociated by pipetting centrifugation at 300g for 5 minutes washed once with PBS and sorted by FACS. Total cell concentration and viability were determined using a TC20 Automated Cell Counter Bio Rad. Samples were then re suspended in 1x PBS with 0.05% BSA at 800 3000 cells per ml. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. Samples were loaded on the 10x Chromium system with a target of 4 000 cells. Chromium Single Cell 3สน GEM Library & Gel Bead Kit v2 or v3 was used for library preparation. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | Oligo-dT | PAIRED | ILLUMINA | Illumina HiSeq 2500 | ERP149744 | Illumina HiSeq 2500 paired end sequencing; Single cell RNA seq 10x Chromium of FACS sorted drl:H2B Dendra2 etv2:Kaede fli1:DsRed and tp1:eGFP transgenic zebrafish embryos | ENA FIRST PUBLIC:2023 08 01|ENA LAST UPDATE:2023 08 01 | drl_h2b-dendra_tb_S33_L001_R2_001.fastq.gz drl_h2b-dendra_tb_S33_L001_R1_001.fastq.gz | fastq fastq | 328918212.0 | 2610462.0 | E MTAB 13196:drl h2b dendra tb S33 L001 | 0:28 1:98 | A:98036464;C:71090137;G:71650807;T:87969481;N:171323 | 28 | 98 | 98036464 | 71090137 | 71650807 | 87969481 | 171323 | ERX11157718 | ERS16172936 | ERA25628220 | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | Institute of Molecular Genetics of the Czech Academy of Sciences|European Nucleotide Archive | 2 | 0.00197 | 0.89969 | 0.00087 | 0.14269 | 0.99681 | 0.89134 | 0.49763 | 0.63651 | 28 | 98 | T | B | sc-like readlen | illumina | hiseq_era | unknown | poly_a | unknown | sc | single_cell_droplet | 10x | Czech Republic | 2023-08-01 | Undetermined | Embryo | Embryo Imprecise | All anatomical structures | ||||||||||||||
| 24656 | 24656 | SRR25491963 | SRX21223193 | SRS18479996 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S12 | GSM7676118 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S12 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676118 | GSM7676118: KBTRE cut replicate S12; Danio rerio; RNA Seq | GSM7676118 r1 | GSM7676118 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTREcut_S12_L001_R2_001.fastq.gz H27K5BCX2_KBTREcut_S12_L001_R1_001.fastq.gz H27K5BCX2_KBTREcut_S12_L001_I1_001.fastq.gz | fastq fastq fastq | 664936140.0 | 5037395.0 | GSM7676118 r1 | 0:8 1:26 2:98 | A:143407233;C:100996340;G:110567290;T:138539687;N:154160 | 8 | 26 | 98 | 143407233 | 100996340 | 110567290 | 138539687 | 154160 | SRX21223193 | SRS18479996 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91249 | 0.11469 | 0.85372 | 0.50388 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24657 | 24657 | SRR25491964 | SRX21223193 | SRS18479996 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S12 | GSM7676118 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S12 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676118 | GSM7676118: KBTRE cut replicate S12; Danio rerio; RNA Seq | GSM7676118 r1 | GSM7676118 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTREcut_S12_L001_R2_001.fastq.gz HFJ5GBCX2_KBTREcut_S12_L001_R1_001.fastq.gz HFJ5GBCX2_KBTREcut_S12_L001_I1_001.fastq.gz | fastq fastq fastq | 645128748.0 | 4887339.0 | GSM7676118 r2 | 0:8 1:26 2:98 | A:139238913;C:98013961;G:107274566;T:134050699;N:381083 | 8 | 26 | 98 | 139238913 | 98013961 | 107274566 | 134050699 | 381083 | SRX21223193 | SRS18479996 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91125 | 0.11373 | 0.85226 | 0.50246 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24658 | 24658 | SRR25491965 | SRX21223193 | SRS18479996 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S12 | GSM7676118 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S12 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676118 | GSM7676118: KBTRE cut replicate S12; Danio rerio; RNA Seq | GSM7676118 r1 | GSM7676118 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTREcut_S12_L001_I1_001.fastq.gz HYLGMBCXY_KBTREcut_S12_L001_R1_001.fastq.gz HYLGMBCXY_KBTREcut_S12_L001_R2_001.fastq.gz | fastq fastq fastq | 701173044.0 | 5311917.0 | GSM7676118 r3 | 0:8 1:26 2:98 | A:151296425;C:106327416;G:116370981;T:144230427;N:2342617 | 8 | 26 | 98 | 151296425 | 106327416 | 116370981 | 144230427 | 2342617 | SRX21223193 | SRS18479996 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91082 | 0.11168 | 0.85245 | 0.5097 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24659 | 24659 | SRR25491966 | SRX21223193 | SRS18479996 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S12 | GSM7676118 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S12 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676118 | GSM7676118: KBTRE cut replicate S12; Danio rerio; RNA Seq | GSM7676118 r1 | GSM7676118 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTREcut_S12_L001_I1_001.fastq.gz HYWGVBCXY_KBTREcut_S12_L001_R1_001.fastq.gz HYWGVBCXY_KBTREcut_S12_L001_R2_001.fastq.gz | fastq fastq fastq | 656204604.0 | 4971247.0 | GSM7676118 r4 | 0:8 1:26 2:98 | A:141756368;C:99522317;G:109015686;T:136785011;N:102824 | 8 | 26 | 98 | 141756368 | 99522317 | 109015686 | 136785011 | 102824 | SRX21223193 | SRS18479996 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.90953 | 0.11383 | 0.85212 | 0.49917 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24660 | 24660 | SRR25492087 | SRX21223193 | SRS18479996 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S12 | GSM7676118 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S12 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676118 | GSM7676118: KBTRE cut replicate S12; Danio rerio; RNA Seq | GSM7676118 r1 | GSM7676118 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTREcut_S12_L002_I1_001.fastq.gz H27K5BCX2_KBTREcut_S12_L002_R1_001.fastq.gz H27K5BCX2_KBTREcut_S12_L002_R2_001.fastq.gz | fastq fastq fastq | 658691616.0 | 4990088.0 | GSM7676118 r5 | 0:8 1:26 2:98 | A:142071317;C:100089239;G:109564812;T:137176685;N:126571 | 8 | 26 | 98 | 142071317 | 100089239 | 109564812 | 137176685 | 126571 | SRX21223193 | SRS18479996 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.9118 | 0.11359 | 0.85307 | 0.49119 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24661 | 24661 | SRR25492088 | SRX21223193 | SRS18479996 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S12 | GSM7676118 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S12 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676118 | GSM7676118: KBTRE cut replicate S12; Danio rerio; RNA Seq | GSM7676118 r1 | GSM7676118 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTREcut_S12_L002_I1_001.fastq.gz HFJ5GBCX2_KBTREcut_S12_L002_R1_001.fastq.gz HFJ5GBCX2_KBTREcut_S12_L002_R2_001.fastq.gz | fastq fastq fastq | 626035872.0 | 4742696.0 | GSM7676118 r6 | 0:8 1:26 2:98 | A:135079818;C:95138940;G:104158101;T:130050700;N:356649 | 8 | 26 | 98 | 135079818 | 95138940 | 104158101 | 130050700 | 356649 | SRX21223193 | SRS18479996 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.9103 | 0.11494 | 0.85378 | 0.48986 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24662 | 24662 | SRR25492089 | SRX21223193 | SRS18479996 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S12 | GSM7676118 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S12 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676118 | GSM7676118: KBTRE cut replicate S12; Danio rerio; RNA Seq | GSM7676118 r1 | GSM7676118 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTREcut_S12_L002_I1_001.fastq.gz HYLGMBCXY_KBTREcut_S12_L002_R1_001.fastq.gz HYLGMBCXY_KBTREcut_S12_L002_R2_001.fastq.gz | fastq fastq fastq | 693479028.0 | 5253629.0 | GSM7676118 r7 | 0:8 1:26 2:98 | A:149676332;C:105134718;G:115079858;T:142696573;N:2268161 | 8 | 26 | 98 | 149676332 | 105134718 | 115079858 | 142696573 | 2268161 | SRX21223193 | SRS18479996 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91107 | 0.11027 | 0.85354 | 0.50421 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24663 | 24663 | SRR25492090 | SRX21223193 | SRS18479996 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S12 | GSM7676118 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S12 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676118 | GSM7676118: KBTRE cut replicate S12; Danio rerio; RNA Seq | GSM7676118 r1 | GSM7676118 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTREcut_S12_L002_R2_001.fastq.gz HYWGVBCXY_KBTREcut_S12_L002_R1_001.fastq.gz HYWGVBCXY_KBTREcut_S12_L002_I1_001.fastq.gz | fastq fastq fastq | 656957664.0 | 4976952.0 | GSM7676118 r8 | 0:8 1:26 2:98 | A:141922523;C:99627529;G:109167065;T:136992486;N:31693 | 8 | 26 | 98 | 141922523 | 99627529 | 109167065 | 136992486 | 31693 | SRX21223193 | SRS18479996 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91107 | 0.11379 | 0.85346 | 0.50695 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24664 | 24664 | SRR25491967 | SRX21223192 | SRS18479995 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S10 | GSM7676116 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S10 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676116 | GSM7676116: KBTRE cut replicate S10; Danio rerio; RNA Seq | GSM7676116 r1 | GSM7676116 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTREcut_S10_L001_R2_001.fastq.gz H27K5BCX2_KBTREcut_S10_L001_R1_001.fastq.gz H27K5BCX2_KBTREcut_S10_L001_I1_001.fastq.gz | fastq fastq fastq | 1062185256.0 | 8046858.0 | GSM7676116 r1 | 0:8 1:26 2:98 | A:230014489;C:161623855;G:176773666;T:219935294;N:244780 | 8 | 26 | 98 | 230014489 | 161623855 | 176773666 | 219935294 | 244780 | SRX21223192 | SRS18479995 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91294 | 0.11493 | 0.85719 | 0.50794 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24665 | 24665 | SRR25491968 | SRX21223192 | SRS18479995 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S10 | GSM7676116 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S10 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676116 | GSM7676116: KBTRE cut replicate S10; Danio rerio; RNA Seq | GSM7676116 r1 | GSM7676116 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTREcut_S10_L001_R2_001.fastq.gz HFJ5GBCX2_KBTREcut_S10_L001_R1_001.fastq.gz HFJ5GBCX2_KBTREcut_S10_L001_I1_001.fastq.gz | fastq fastq fastq | 1034539308.0 | 7837419.0 | GSM7676116 r2 | 0:8 1:26 2:98 | A:224616171;C:157304961;G:172084243;T:213455656;N:606031 | 8 | 26 | 98 | 224616171 | 157304961 | 172084243 | 213455656 | 606031 | SRX21223192 | SRS18479995 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91287 | 0.11588 | 0.85415 | 0.50804 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24666 | 24666 | SRR25491969 | SRX21223192 | SRS18479995 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S10 | GSM7676116 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S10 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676116 | GSM7676116: KBTRE cut replicate S10; Danio rerio; RNA Seq | GSM7676116 r1 | GSM7676116 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTREcut_S10_L001_R2_001.fastq.gz HYLGMBCXY_KBTREcut_S10_L001_R1_001.fastq.gz HYLGMBCXY_KBTREcut_S10_L001_I1_001.fastq.gz | fastq fastq fastq | 1122858660.0 | 8506505.0 | GSM7676116 r3 | 0:8 1:26 2:98 | A:243355100;C:170483208;G:186449194;T:229601473;N:3748515 | 8 | 26 | 98 | 243355100 | 170483208 | 186449194 | 229601473 | 3748515 | SRX21223192 | SRS18479995 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91213 | 0.11262 | 0.8561 | 0.49742 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24667 | 24667 | SRR25491970 | SRX21223192 | SRS18479995 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S10 | GSM7676116 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S10 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676116 | GSM7676116: KBTRE cut replicate S10; Danio rerio; RNA Seq | GSM7676116 r1 | GSM7676116 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTREcut_S10_L001_I1_001.fastq.gz HYWGVBCXY_KBTREcut_S10_L001_R1_001.fastq.gz HYWGVBCXY_KBTREcut_S10_L001_R2_001.fastq.gz | fastq fastq fastq | 1044506496.0 | 7912928.0 | GSM7676116 r4 | 0:8 1:26 2:98 | A:226413586;C:158757664;G:173768785;T:216364905;N:162004 | 8 | 26 | 98 | 226413586 | 158757664 | 173768785 | 216364905 | 162004 | SRX21223192 | SRS18479995 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.9121 | 0.11332 | 0.85401 | 0.51062 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24668 | 24668 | SRR25491971 | SRX21223192 | SRS18479995 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S10 | GSM7676116 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S10 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676116 | GSM7676116: KBTRE cut replicate S10; Danio rerio; RNA Seq | GSM7676116 r1 | GSM7676116 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTREcut_S10_L002_I1_001.fastq.gz HYLGMBCXY_KBTREcut_S10_L002_R1_001.fastq.gz HYLGMBCXY_KBTREcut_S10_L002_R2_001.fastq.gz | fastq fastq fastq | 1110246720.0 | 8410960.0 | GSM7676116 r7 | 0:8 1:26 2:98 | A:240787456;C:168521971;G:184254856;T:227029149;N:3680648 | 8 | 26 | 98 | 240787456 | 168521971 | 184254856 | 227029149 | 3680648 | SRX21223192 | SRS18479995 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91246 | 0.11301 | 0.85449 | 0.50422 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24669 | 24669 | SRR25491972 | SRX21223192 | SRS18479995 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S10 | GSM7676116 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S10 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676116 | GSM7676116: KBTRE cut replicate S10; Danio rerio; RNA Seq | GSM7676116 r1 | GSM7676116 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTREcut_S10_L002_I1_001.fastq.gz HYWGVBCXY_KBTREcut_S10_L002_R1_001.fastq.gz HYWGVBCXY_KBTREcut_S10_L002_R2_001.fastq.gz | fastq fastq fastq | 1046026740.0 | 7924445.0 | GSM7676116 r8 | 0:8 1:26 2:98 | A:226911006;C:158976256;G:173939936;T:216718492;N:49920 | 8 | 26 | 98 | 226911006 | 158976256 | 173939936 | 216718492 | 49920 | SRX21223192 | SRS18479995 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91267 | 0.11377 | 0.8548 | 0.50065 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24670 | 24670 | SRR25491989 | SRX21223192 | SRS18479995 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S10 | GSM7676116 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S10 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676116 | GSM7676116: KBTRE cut replicate S10; Danio rerio; RNA Seq | GSM7676116 r1 | GSM7676116 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTREcut_S10_L002_I1_001.fastq.gz H27K5BCX2_KBTREcut_S10_L002_R1_001.fastq.gz H27K5BCX2_KBTREcut_S10_L002_R2_001.fastq.gz | fastq fastq fastq | 1052457252.0 | 7973161.0 | GSM7676116 r5 | 0:8 1:26 2:98 | A:227727118;C:160159228;G:175278732;T:218004542;N:200158 | 8 | 26 | 98 | 227727118 | 160159228 | 175278732 | 218004542 | 200158 | SRX21223192 | SRS18479995 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91307 | 0.11551 | 0.85634 | 0.50392 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24671 | 24671 | SRR25491990 | SRX21223192 | SRS18479995 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S10 | GSM7676116 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S10 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676116 | GSM7676116: KBTRE cut replicate S10; Danio rerio; RNA Seq | GSM7676116 r1 | GSM7676116 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTREcut_S10_L002_I1_001.fastq.gz HFJ5GBCX2_KBTREcut_S10_L002_R1_001.fastq.gz HFJ5GBCX2_KBTREcut_S10_L002_R2_001.fastq.gz | fastq fastq fastq | 1003744500.0 | 7604125.0 | GSM7676116 r6 | 0:8 1:26 2:98 | A:217741909;C:152712224;G:167110758;T:207073522;N:565837 | 8 | 26 | 98 | 217741909 | 152712224 | 167110758 | 207073522 | 565837 | SRX21223192 | SRS18479995 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91357 | 0.11506 | 0.85504 | 0.49763 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24672 | 24672 | SRR25491973 | SRX21223191 | SRS18479994 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S11 | GSM7676117 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S11 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676117 | GSM7676117: KBTRE cut replicate S11; Danio rerio; RNA Seq | GSM7676117 r1 | GSM7676117 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTREcut_S11_L001_I1_001.fastq.gz H27K5BCX2_KBTREcut_S11_L001_R1_001.fastq.gz H27K5BCX2_KBTREcut_S11_L001_R2_001.fastq.gz | fastq fastq fastq | 927688740.0 | 7027945.0 | GSM7676117 r1 | 0:8 1:26 2:98 | A:200105466;C:141029173;G:154321296;T:193068023;N:214652 | 8 | 26 | 98 | 200105466 | 141029173 | 154321296 | 193068023 | 214652 | SRX21223191 | SRS18479994 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91282 | 0.1155 | 0.85283 | 0.5061 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24673 | 24673 | SRR25491974 | SRX21223191 | SRS18479994 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S11 | GSM7676117 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S11 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676117 | GSM7676117: KBTRE cut replicate S11; Danio rerio; RNA Seq | GSM7676117 r1 | GSM7676117 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTREcut_S11_L001_R2_001.fastq.gz HFJ5GBCX2_KBTREcut_S11_L001_R1_001.fastq.gz HFJ5GBCX2_KBTREcut_S11_L001_I1_001.fastq.gz | fastq fastq fastq | 897789420.0 | 6801435.0 | GSM7676117 r2 | 0:8 1:26 2:98 | A:193853631;C:136435624;G:149320255;T:186400739;N:530381 | 8 | 26 | 98 | 193853631 | 136435624 | 149320255 | 186400739 | 530381 | SRX21223191 | SRS18479994 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91186 | 0.11485 | 0.85429 | 0.49592 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24674 | 24674 | SRR25491975 | SRX21223191 | SRS18479994 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S11 | GSM7676117 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S11 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676117 | GSM7676117: KBTRE cut replicate S11; Danio rerio; RNA Seq | GSM7676117 r1 | GSM7676117 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTREcut_S11_L001_R2_001.fastq.gz HYLGMBCXY_KBTREcut_S11_L001_R1_001.fastq.gz HYLGMBCXY_KBTREcut_S11_L001_I1_001.fastq.gz | fastq fastq fastq | 978294372.0 | 7411321.0 | GSM7676117 r3 | 0:8 1:26 2:98 | A:211132117;C:148425540;G:162317813;T:201164320;N:3269668 | 8 | 26 | 98 | 211132117 | 148425540 | 162317813 | 201164320 | 3269668 | SRX21223191 | SRS18479994 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91073 | 0.1132 | 0.8537 | 0.50609 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24675 | 24675 | SRR25491976 | SRX21223191 | SRS18479994 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S11 | GSM7676117 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S11 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676117 | GSM7676117: KBTRE cut replicate S11; Danio rerio; RNA Seq | GSM7676117 r1 | GSM7676117 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTREcut_S11_L001_I1_001.fastq.gz HYWGVBCXY_KBTREcut_S11_L001_R1_001.fastq.gz HYWGVBCXY_KBTREcut_S11_L001_R2_001.fastq.gz | fastq fastq fastq | 918264996.0 | 6956553.0 | GSM7676117 r4 | 0:8 1:26 2:98 | A:198352554;C:139394187;G:152576773;T:191275437;N:143243 | 8 | 26 | 98 | 198352554 | 139394187 | 152576773 | 191275437 | 143243 | SRX21223191 | SRS18479994 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91014 | 0.1139 | 0.85307 | 0.50536 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24676 | 24676 | SRR25491977 | SRX21223191 | SRS18479994 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S11 | GSM7676117 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S11 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676117 | GSM7676117: KBTRE cut replicate S11; Danio rerio; RNA Seq | GSM7676117 r1 | GSM7676117 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTREcut_S11_L002_I1_001.fastq.gz H27K5BCX2_KBTREcut_S11_L002_R1_001.fastq.gz H27K5BCX2_KBTREcut_S11_L002_R2_001.fastq.gz | fastq fastq fastq | 918134580.0 | 6955565.0 | GSM7676117 r5 | 0:8 1:26 2:98 | A:198094257;C:139568875;G:152735838;T:191072590;N:173810 | 8 | 26 | 98 | 198094257 | 139568875 | 152735838 | 191072590 | 173810 | SRX21223191 | SRS18479994 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91196 | 0.11403 | 0.85267 | 0.50361 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24677 | 24677 | SRR25491978 | SRX21223191 | SRS18479994 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S11 | GSM7676117 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S11 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676117 | GSM7676117: KBTRE cut replicate S11; Danio rerio; RNA Seq | GSM7676117 r1 | GSM7676117 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTREcut_S11_L002_I1_001.fastq.gz HFJ5GBCX2_KBTREcut_S11_L002_R1_001.fastq.gz HFJ5GBCX2_KBTREcut_S11_L002_R2_001.fastq.gz | fastq fastq fastq | 868525152.0 | 6579736.0 | GSM7676117 r6 | 0:8 1:26 2:98 | A:187572958;C:132014722;G:144495273;T:180237409;N:493766 | 8 | 26 | 98 | 187572958 | 132014722 | 144495273 | 180237409 | 493766 | SRX21223191 | SRS18479994 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91049 | 0.11265 | 0.85273 | 0.49392 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24678 | 24678 | SRR25491979 | SRX21223191 | SRS18479994 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S11 | GSM7676117 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S11 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676117 | GSM7676117: KBTRE cut replicate S11; Danio rerio; RNA Seq | GSM7676117 r1 | GSM7676117 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTREcut_S11_L002_I1_001.fastq.gz HYLGMBCXY_KBTREcut_S11_L002_R1_001.fastq.gz HYLGMBCXY_KBTREcut_S11_L002_R2_001.fastq.gz | fastq fastq fastq | 967222872.0 | 7327446.0 | GSM7676117 r7 | 0:8 1:26 2:98 | A:208757589;C:146699883;G:160504752;T:198933585;N:3193899 | 8 | 26 | 98 | 208757589 | 146699883 | 160504752 | 198933585 | 3193899 | SRX21223191 | SRS18479994 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91068 | 0.11108 | 0.85449 | 0.48271 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24679 | 24679 | SRR25491980 | SRX21223191 | SRS18479994 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE cut replicate S11 | GSM7676117 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE cut replicate S11 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676117 | GSM7676117: KBTRE cut replicate S11; Danio rerio; RNA Seq | GSM7676117 r1 | GSM7676117 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTREcut_S11_L002_I1_001.fastq.gz HYWGVBCXY_KBTREcut_S11_L002_R1_001.fastq.gz HYWGVBCXY_KBTREcut_S11_L002_R2_001.fastq.gz | fastq fastq fastq | 918366636.0 | 6957323.0 | GSM7676117 r8 | 0:8 1:26 2:98 | A:198420992;C:139446089;G:152636322;T:191270874;N:43377 | 8 | 26 | 98 | 198420992 | 139446089 | 152636322 | 191270874 | 43377 | SRX21223191 | SRS18479994 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.91125 | 0.11279 | 0.85212 | 0.49708 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24680 | 24680 | SRR25491981 | SRX21223190 | SRS18479993 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S8 | GSM7676115 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S8 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676115 | GSM7676115: KBTGR normal replicate S8; Danio rerio; RNA Seq | GSM7676115 r1 | GSM7676115 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTGRnormal_S8_L001_I1_001.fastq.gz H27K5BCX2_KBTGRnormal_S8_L001_R1_001.fastq.gz H27K5BCX2_KBTGRnormal_S8_L001_R2_001.fastq.gz | fastq fastq fastq | 1367764332.0 | 10361851.0 | GSM7676115 r1 | 0:8 1:26 2:98 | A:291792120;C:211871917;G:233940380;T:277543215;N:313766 | 8 | 26 | 98 | 291792120 | 211871917 | 233940380 | 277543215 | 313766 | SRX21223190 | SRS18479993 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92841 | 0.09159 | 0.82191 | 0.48306 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24681 | 24681 | SRR25491982 | SRX21223190 | SRS18479993 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S8 | GSM7676115 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S8 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676115 | GSM7676115: KBTGR normal replicate S8; Danio rerio; RNA Seq | GSM7676115 r1 | GSM7676115 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTGRnormal_S8_L001_I1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S8_L001_R1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S8_L001_R2_001.fastq.gz | fastq fastq fastq | 1324750284.0 | 10035987.0 | GSM7676115 r2 | 0:8 1:26 2:98 | A:282670428;C:205193043;G:226649784;T:268220709;N:792762 | 8 | 26 | 98 | 282670428 | 205193043 | 226649784 | 268220709 | 792762 | SRX21223190 | SRS18479993 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92859 | 0.09205 | 0.82039 | 0.48039 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24682 | 24682 | SRR25491983 | SRX21223190 | SRS18479993 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S8 | GSM7676115 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S8 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676115 | GSM7676115: KBTGR normal replicate S8; Danio rerio; RNA Seq | GSM7676115 r1 | GSM7676115 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTGRnormal_S8_L001_I1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S8_L001_R1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S8_L001_R2_001.fastq.gz | fastq fastq fastq | 1442154120.0 | 10925410.0 | GSM7676115 r3 | 0:8 1:26 2:98 | A:307172615;C:223150292;G:246311885;T:289234193;N:4821195 | 8 | 26 | 98 | 307172615 | 223150292 | 246311885 | 289234193 | 4821195 | SRX21223190 | SRS18479993 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92841 | 0.08941 | 0.82331 | 0.49215 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24683 | 24683 | SRR25491984 | SRX21223190 | SRS18479993 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S8 | GSM7676115 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S8 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676115 | GSM7676115: KBTGR normal replicate S8; Danio rerio; RNA Seq | GSM7676115 r1 | GSM7676115 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTGRnormal_S8_L001_I1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S8_L001_R1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S8_L001_R2_001.fastq.gz | fastq fastq fastq | 1358634684.0 | 10292687.0 | GSM7676115 r4 | 0:8 1:26 2:98 | A:290016958;C:210290529;G:232317191;T:275847772;N:210876 | 8 | 26 | 98 | 290016958 | 210290529 | 232317191 | 275847772 | 210876 | SRX21223190 | SRS18479993 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92745 | 0.09192 | 0.82229 | 0.49666 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24684 | 24684 | SRR25491985 | SRX21223190 | SRS18479993 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S8 | GSM7676115 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S8 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676115 | GSM7676115: KBTGR normal replicate S8; Danio rerio; RNA Seq | GSM7676115 r1 | GSM7676115 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTGRnormal_S8_L002_I1_001.fastq.gz H27K5BCX2_KBTGRnormal_S8_L002_R1_001.fastq.gz H27K5BCX2_KBTGRnormal_S8_L002_R2_001.fastq.gz | fastq fastq fastq | 1353664092.0 | 10255031.0 | GSM7676115 r5 | 0:8 1:26 2:98 | A:288847376;C:209739068;G:231593033;T:274552362;N:261199 | 8 | 26 | 98 | 288847376 | 209739068 | 231593033 | 274552362 | 261199 | SRX21223190 | SRS18479993 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92838 | 0.09211 | 0.82207 | 0.48301 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24685 | 24685 | SRR25491986 | SRX21223190 | SRS18479993 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S8 | GSM7676115 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S8 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676115 | GSM7676115: KBTGR normal replicate S8; Danio rerio; RNA Seq | GSM7676115 r1 | GSM7676115 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTGRnormal_S8_L002_I1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S8_L002_R1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S8_L002_R2_001.fastq.gz | fastq fastq fastq | 1282964496.0 | 9719428.0 | GSM7676115 r6 | 0:8 1:26 2:98 | A:273803342;C:198769671;G:219587842;T:259615174;N:727915 | 8 | 26 | 98 | 273803342 | 198769671 | 219587842 | 259615174 | 727915 | SRX21223190 | SRS18479993 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92815 | 0.09094 | 0.82189 | 0.47933 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24686 | 24686 | SRR25491987 | SRX21223190 | SRS18479993 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S8 | GSM7676115 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S8 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676115 | GSM7676115: KBTGR normal replicate S8; Danio rerio; RNA Seq | GSM7676115 r1 | GSM7676115 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTGRnormal_S8_L002_I1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S8_L002_R1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S8_L002_R2_001.fastq.gz | fastq fastq fastq | 1425314880.0 | 10797840.0 | GSM7676115 r7 | 0:8 1:26 2:98 | A:303654013;C:220453444;G:243409971;T:285937241;N:4733651 | 8 | 26 | 98 | 303654013 | 220453444 | 243409971 | 285937241 | 4733651 | SRX21223190 | SRS18479993 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92724 | 0.09059 | 0.82345 | 0.49626 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24687 | 24687 | SRR25491988 | SRX21223190 | SRS18479993 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S8 | GSM7676115 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S8 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676115 | GSM7676115: KBTGR normal replicate S8; Danio rerio; RNA Seq | GSM7676115 r1 | GSM7676115 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTGRnormal_S8_L002_I1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S8_L002_R1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S8_L002_R2_001.fastq.gz | fastq fastq fastq | 1358584788.0 | 10292309.0 | GSM7676115 r8 | 0:8 1:26 2:98 | A:290131069;C:210271668;G:232237599;T:275940105;N:65841 | 8 | 26 | 98 | 290131069 | 210271668 | 232237599 | 275940105 | 65841 | SRX21223190 | SRS18479993 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92858 | 0.09144 | 0.82089 | 0.49057 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24688 | 24688 | SRR25491991 | SRX21223189 | SRS18479992 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S7 | GSM7676114 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S7 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676114 | GSM7676114: KBTGR normal replicate S7; Danio rerio; RNA Seq | GSM7676114 r1 | GSM7676114 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTGRnormal_S7_L001_I1_001.fastq.gz H27K5BCX2_KBTGRnormal_S7_L001_R1_001.fastq.gz H27K5BCX2_KBTGRnormal_S7_L001_R2_001.fastq.gz | fastq fastq fastq | 1045774224.0 | 7922532.0 | GSM7676114 r1 | 0:8 1:26 2:98 | A:224743708;C:161896286;G:178467180;T:211061243;N:239719 | 8 | 26 | 98 | 224743708 | 161896286 | 178467180 | 211061243 | 239719 | SRX21223189 | SRS18479992 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92898 | 0.0948 | 0.82576 | 0.48767 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24689 | 24689 | SRR25491992 | SRX21223189 | SRS18479992 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S7 | GSM7676114 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S7 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676114 | GSM7676114: KBTGR normal replicate S7; Danio rerio; RNA Seq | GSM7676114 r1 | GSM7676114 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTGRnormal_S7_L001_I1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S7_L001_R1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S7_L001_R2_001.fastq.gz | fastq fastq fastq | 1023737880.0 | 7755590.0 | GSM7676114 r2 | 0:8 1:26 2:98 | A:220339751;C:158506697;G:174717320;T:205879340;N:604712 | 8 | 26 | 98 | 220339751 | 158506697 | 174717320 | 205879340 | 604712 | SRX21223189 | SRS18479992 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.93138 | 0.09445 | 0.82319 | 0.49095 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24690 | 24690 | SRR25491993 | SRX21223189 | SRS18479992 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S7 | GSM7676114 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S7 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676114 | GSM7676114: KBTGR normal replicate S7; Danio rerio; RNA Seq | GSM7676114 r1 | GSM7676114 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTGRnormal_S7_L001_I1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S7_L001_R1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S7_L001_R2_001.fastq.gz | fastq fastq fastq | 1107722484.0 | 8391837.0 | GSM7676114 r3 | 0:8 1:26 2:98 | A:237628383;C:171310823;G:188847113;T:220892571;N:3721136 | 8 | 26 | 98 | 237628383 | 171310823 | 188847113 | 220892571 | 3721136 | SRX21223189 | SRS18479992 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92932 | 0.09165 | 0.82432 | 0.48843 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24691 | 24691 | SRR25491994 | SRX21223189 | SRS18479992 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S7 | GSM7676114 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S7 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676114 | GSM7676114: KBTGR normal replicate S7; Danio rerio; RNA Seq | GSM7676114 r1 | GSM7676114 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTGRnormal_S7_L001_I1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S7_L001_R1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S7_L001_R2_001.fastq.gz | fastq fastq fastq | 1018155600.0 | 7713300.0 | GSM7676114 r4 | 0:8 1:26 2:98 | A:218785960;C:157467573;G:173782025;T:205710180;N:157662 | 8 | 26 | 98 | 218785960 | 157467573 | 173782025 | 205710180 | 157662 | SRX21223189 | SRS18479992 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92735 | 0.09294 | 0.82304 | 0.48936 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24692 | 24692 | SRR25491995 | SRX21223189 | SRS18479992 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S7 | GSM7676114 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S7 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676114 | GSM7676114: KBTGR normal replicate S7; Danio rerio; RNA Seq | GSM7676114 r1 | GSM7676114 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTGRnormal_S7_L002_I1_001.fastq.gz H27K5BCX2_KBTGRnormal_S7_L002_R1_001.fastq.gz H27K5BCX2_KBTGRnormal_S7_L002_R2_001.fastq.gz | fastq fastq fastq | 1037896992.0 | 7862856.0 | GSM7676114 r5 | 0:8 1:26 2:98 | A:222624793;C:160770752;G:177337819;T:209629234;N:197290 | 8 | 26 | 98 | 222624793 | 160770752 | 177337819 | 209629234 | 197290 | SRX21223189 | SRS18479992 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.93038 | 0.09291 | 0.82513 | 0.48834 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24693 | 24693 | SRR25491996 | SRX21223189 | SRS18479992 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S7 | GSM7676114 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S7 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676114 | GSM7676114: KBTGR normal replicate S7; Danio rerio; RNA Seq | GSM7676114 r1 | GSM7676114 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTGRnormal_S7_L002_I1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S7_L002_R1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S7_L002_R2_001.fastq.gz | fastq fastq fastq | 994648116.0 | 7535213.0 | GSM7676114 r6 | 0:8 1:26 2:98 | A:213918528;C:154046950;G:169913173;T:200006956;N:565267 | 8 | 26 | 98 | 213918528 | 154046950 | 169913173 | 200006956 | 565267 | SRX21223189 | SRS18479992 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92863 | 0.09237 | 0.8244 | 0.48574 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24694 | 24694 | SRR25491997 | SRX21223189 | SRS18479992 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S7 | GSM7676114 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S7 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676114 | GSM7676114: KBTGR normal replicate S7; Danio rerio; RNA Seq | GSM7676114 r1 | GSM7676114 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTGRnormal_S7_L002_I1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S7_L002_R1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S7_L002_R2_001.fastq.gz | fastq fastq fastq | 1095910068.0 | 8302349.0 | GSM7676114 r7 | 0:8 1:26 2:98 | A:235361030;C:169424678;G:186752468;T:218466797;N:3625229 | 8 | 26 | 98 | 235361030 | 169424678 | 186752468 | 218466797 | 3625229 | SRX21223189 | SRS18479992 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92887 | 0.09019 | 0.82696 | 0.4934 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24695 | 24695 | SRR25491998 | SRX21223189 | SRS18479992 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S7 | GSM7676114 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S7 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676114 | GSM7676114: KBTGR normal replicate S7; Danio rerio; RNA Seq | GSM7676114 r1 | GSM7676114 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTGRnormal_S7_L002_I1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S7_L002_R1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S7_L002_R2_001.fastq.gz | fastq fastq fastq | 1020164640.0 | 7728520.0 | GSM7676114 r8 | 0:8 1:26 2:98 | A:219455597;C:157669416;G:174033845;T:206187545;N:48557 | 8 | 26 | 98 | 219455597 | 157669416 | 174033845 | 206187545 | 48557 | SRX21223189 | SRS18479992 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.93 | 0.0929 | 0.82507 | 0.48822 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24696 | 24696 | SRR25491999 | SRX21223188 | SRS18479991 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S6 | GSM7676113 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S6 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676113 | GSM7676113: KBTGR normal replicate S6; Danio rerio; RNA Seq | GSM7676113 r1 | GSM7676113 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTGRnormal_S6_L001_I1_001.fastq.gz H27K5BCX2_KBTGRnormal_S6_L001_R1_001.fastq.gz H27K5BCX2_KBTGRnormal_S6_L001_R2_001.fastq.gz | fastq fastq fastq | 1289409660.0 | 9768255.0 | GSM7676113 r1 | 0:8 1:26 2:98 | A:275297633;C:199461255;G:220403674;T:261830960;N:295468 | 8 | 26 | 98 | 275297633 | 199461255 | 220403674 | 261830960 | 295468 | SRX21223188 | SRS18479991 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92761 | 0.09178 | 0.82089 | 0.49952 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24697 | 24697 | SRR25492000 | SRX21223188 | SRS18479991 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S6 | GSM7676113 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S6 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676113 | GSM7676113: KBTGR normal replicate S6; Danio rerio; RNA Seq | GSM7676113 r1 | GSM7676113 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTGRnormal_S6_L001_I1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S6_L001_R1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S6_L001_R2_001.fastq.gz | fastq fastq fastq | 1254104148.0 | 9500789.0 | GSM7676113 r2 | 0:8 1:26 2:98 | A:267759276;C:194047461;G:214469796;T:254057804;N:742985 | 8 | 26 | 98 | 267759276 | 194047461 | 214469796 | 254057804 | 742985 | SRX21223188 | SRS18479991 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92863 | 0.09263 | 0.82016 | 0.49849 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24698 | 24698 | SRR25492001 | SRX21223188 | SRS18479991 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S6 | GSM7676113 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S6 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676113 | GSM7676113: KBTGR normal replicate S6; Danio rerio; RNA Seq | GSM7676113 r1 | GSM7676113 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTGRnormal_S6_L001_I1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S6_L001_R1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S6_L001_R2_001.fastq.gz | fastq fastq fastq | 1364037444.0 | 10333617.0 | GSM7676113 r3 | 0:8 1:26 2:98 | A:290782963;C:210764004;G:232828321;T:273741440;N:4577738 | 8 | 26 | 98 | 290782963 | 210764004 | 232828321 | 273741440 | 4577738 | SRX21223188 | SRS18479991 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92727 | 0.09037 | 0.82339 | 0.49743 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24699 | 24699 | SRR25492002 | SRX21223188 | SRS18479991 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S6 | GSM7676113 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S6 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676113 | GSM7676113: KBTGR normal replicate S6; Danio rerio; RNA Seq | GSM7676113 r1 | GSM7676113 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTGRnormal_S6_L001_I1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S6_L001_R1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S6_L001_R2_001.fastq.gz | fastq fastq fastq | 1274535900.0 | 9655575.0 | GSM7676113 r4 | 0:8 1:26 2:98 | A:272331035;C:196919570;G:217674747;T:259126322;N:194676 | 8 | 26 | 98 | 272331035 | 196919570 | 217674747 | 259126322 | 194676 | SRX21223188 | SRS18479991 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.9267 | 0.09131 | 0.82193 | 0.49736 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24700 | 24700 | SRR25492003 | SRX21223188 | SRS18479991 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S6 | GSM7676113 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S6 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676113 | GSM7676113: KBTGR normal replicate S6; Danio rerio; RNA Seq | GSM7676113 r1 | GSM7676113 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTGRnormal_S6_L002_I1_001.fastq.gz H27K5BCX2_KBTGRnormal_S6_L002_R1_001.fastq.gz H27K5BCX2_KBTGRnormal_S6_L002_R2_001.fastq.gz | fastq fastq fastq | 1278788544.0 | 9687792.0 | GSM7676113 r5 | 0:8 1:26 2:98 | A:273089526;C:197794682;G:218629305;T:259640660;N:249443 | 8 | 26 | 98 | 273089526 | 197794682 | 218629305 | 259640660 | 249443 | SRX21223188 | SRS18479991 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92837 | 0.09211 | 0.82069 | 0.49043 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24701 | 24701 | SRR25492004 | SRX21223188 | SRS18479991 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S6 | GSM7676113 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S6 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676113 | GSM7676113: KBTGR normal replicate S6; Danio rerio; RNA Seq | GSM7676113 r1 | GSM7676113 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTGRnormal_S6_L002_I1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S6_L002_R1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S6_L002_R2_001.fastq.gz | fastq fastq fastq | 1218538200.0 | 9231350.0 | GSM7676113 r6 | 0:8 1:26 2:98 | A:260174350;C:188588952;G:208442781;T:246771528;N:694689 | 8 | 26 | 98 | 260174350 | 188588952 | 208442781 | 246771528 | 694689 | SRX21223188 | SRS18479991 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92755 | 0.09174 | 0.82152 | 0.48524 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24702 | 24702 | SRR25492005 | SRX21223188 | SRS18479991 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S6 | GSM7676113 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S6 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676113 | GSM7676113: KBTGR normal replicate S6; Danio rerio; RNA Seq | GSM7676113 r1 | GSM7676113 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTGRnormal_S6_L002_I1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S6_L002_R1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S6_L002_R2_001.fastq.gz | fastq fastq fastq | 1348885428.0 | 10218829.0 | GSM7676113 r7 | 0:8 1:26 2:98 | A:287599265;C:208361077;G:230218011;T:270815023;N:4451866 | 8 | 26 | 98 | 287599265 | 208361077 | 230218011 | 270815023 | 4451866 | SRX21223188 | SRS18479991 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92616 | 0.08938 | 0.82189 | 0.49517 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24703 | 24703 | SRR25492006 | SRX21223188 | SRS18479991 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S6 | GSM7676113 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S6 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676113 | GSM7676113: KBTGR normal replicate S6; Danio rerio; RNA Seq | GSM7676113 r1 | GSM7676113 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTGRnormal_S6_L002_I1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S6_L002_R1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S6_L002_R2_001.fastq.gz | fastq fastq fastq | 1274044596.0 | 9651853.0 | GSM7676113 r8 | 0:8 1:26 2:98 | A:272274110;C:196868273;G:217614843;T:259062639;N:61729 | 8 | 26 | 98 | 272274110 | 196868273 | 217614843 | 259062639 | 61729 | SRX21223188 | SRS18479991 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92673 | 0.09105 | 0.82142 | 0.48867 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24704 | 24704 | SRR25492007 | SRX21223187 | SRS18479990 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S5 | GSM7676112 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S5 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676112 | GSM7676112: KBTGR normal replicate S5; Danio rerio; RNA Seq | GSM7676112 r1 | GSM7676112 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTGRnormal_S5_L001_I1_001.fastq.gz H27K5BCX2_KBTGRnormal_S5_L001_R1_001.fastq.gz H27K5BCX2_KBTGRnormal_S5_L001_R2_001.fastq.gz | fastq fastq fastq | 1258754244.0 | 9536017.0 | GSM7676112 r1 | 0:8 1:26 2:98 | A:269418553;C:194370105;G:214652075;T:255806804;N:282129 | 8 | 26 | 98 | 269418553 | 194370105 | 214652075 | 255806804 | 282129 | SRX21223187 | SRS18479990 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.9286 | 0.09296 | 0.82288 | 0.48038 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24705 | 24705 | SRR25492008 | SRX21223187 | SRS18479990 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S5 | GSM7676112 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S5 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676112 | GSM7676112: KBTGR normal replicate S5; Danio rerio; RNA Seq | GSM7676112 r1 | GSM7676112 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTGRnormal_S5_L001_I1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S5_L001_R1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S5_L001_R2_001.fastq.gz | fastq fastq fastq | 1217159724.0 | 9220907.0 | GSM7676112 r2 | 0:8 1:26 2:98 | A:260789467;C:187945962;G:207550436;T:246642449;N:720572 | 8 | 26 | 98 | 260789467 | 187945962 | 207550436 | 246642449 | 720572 | SRX21223187 | SRS18479990 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.93045 | 0.09356 | 0.82386 | 0.4736 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24706 | 24706 | SRR25492009 | SRX21223187 | SRS18479990 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S5 | GSM7676112 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S5 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676112 | GSM7676112: KBTGR normal replicate S5; Danio rerio; RNA Seq | GSM7676112 r1 | GSM7676112 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTGRnormal_S5_L001_I1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S5_L001_R1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S5_L001_R2_001.fastq.gz | fastq fastq fastq | 1327996824.0 | 10060582.0 | GSM7676112 r3 | 0:8 1:26 2:98 | A:283722392;C:204894097;G:226054109;T:266823827;N:4442611 | 8 | 26 | 98 | 283722392 | 204894097 | 226054109 | 266823827 | 4442611 | SRX21223187 | SRS18479990 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92793 | 0.09118 | 0.82513 | 0.48007 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24707 | 24707 | SRR25492010 | SRX21223187 | SRS18479990 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S5 | GSM7676112 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S5 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676112 | GSM7676112: KBTGR normal replicate S5; Danio rerio; RNA Seq | GSM7676112 r1 | GSM7676112 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTGRnormal_S5_L001_I1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S5_L001_R1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S5_L001_R2_001.fastq.gz | fastq fastq fastq | 1255655808.0 | 9512544.0 | GSM7676112 r4 | 0:8 1:26 2:98 | A:268848025;C:193752306;G:214061624;T:255371933;N:195424 | 8 | 26 | 98 | 268848025 | 193752306 | 214061624 | 255371933 | 195424 | SRX21223187 | SRS18479990 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92719 | 0.09327 | 0.82221 | 0.47626 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24708 | 24708 | SRR25492011 | SRX21223187 | SRS18479990 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S5 | GSM7676112 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S5 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676112 | GSM7676112: KBTGR normal replicate S5; Danio rerio; RNA Seq | GSM7676112 r1 | GSM7676112 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTGRnormal_S5_L002_I1_001.fastq.gz H27K5BCX2_KBTGRnormal_S5_L002_R1_001.fastq.gz H27K5BCX2_KBTGRnormal_S5_L002_R2_001.fastq.gz | fastq fastq fastq | 1246449072.0 | 9442796.0 | GSM7676112 r5 | 0:8 1:26 2:98 | A:266772235;C:192514744;G:212592908;T:253285733;N:228388 | 8 | 26 | 98 | 266772235 | 192514744 | 212592908 | 253285733 | 228388 | SRX21223187 | SRS18479990 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92776 | 0.09295 | 0.82333 | 0.48441 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24709 | 24709 | SRR25492012 | SRX21223187 | SRS18479990 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S5 | GSM7676112 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S5 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676112 | GSM7676112: KBTGR normal replicate S5; Danio rerio; RNA Seq | GSM7676112 r1 | GSM7676112 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTGRnormal_S5_L002_I1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S5_L002_R1_001.fastq.gz HFJ5GBCX2_KBTGRnormal_S5_L002_R2_001.fastq.gz | fastq fastq fastq | 1176943812.0 | 8916241.0 | GSM7676112 r6 | 0:8 1:26 2:98 | A:252114687;C:181782204;G:200782194;T:238444957;N:667576 | 8 | 26 | 98 | 252114687 | 181782204 | 200782194 | 238444957 | 667576 | SRX21223187 | SRS18479990 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92798 | 0.09407 | 0.82225 | 0.47898 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24710 | 24710 | SRR25492013 | SRX21223187 | SRS18479990 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S5 | GSM7676112 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S5 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676112 | GSM7676112: KBTGR normal replicate S5; Danio rerio; RNA Seq | GSM7676112 r1 | GSM7676112 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTGRnormal_S5_L002_I1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S5_L002_R1_001.fastq.gz HYLGMBCXY_KBTGRnormal_S5_L002_R2_001.fastq.gz | fastq fastq fastq | 1312162368.0 | 9940624.0 | GSM7676112 r7 | 0:8 1:26 2:98 | A:280415120;C:202362829;G:223408996;T:263680714;N:4313493 | 8 | 26 | 98 | 280415120 | 202362829 | 223408996 | 263680714 | 4313493 | SRX21223187 | SRS18479990 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.9269 | 0.09153 | 0.82386 | 0.48775 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24711 | 24711 | SRR25492014 | SRX21223187 | SRS18479990 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR normal replicate S5 | GSM7676112 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR normal replicate S5 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676112 | GSM7676112: KBTGR normal replicate S5; Danio rerio; RNA Seq | GSM7676112 r1 | GSM7676112 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTGRnormal_S5_L002_I1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S5_L002_R1_001.fastq.gz HYWGVBCXY_KBTGRnormal_S5_L002_R2_001.fastq.gz | fastq fastq fastq | 1254798732.0 | 9506051.0 | GSM7676112 r8 | 0:8 1:26 2:98 | A:268758608;C:193670297;G:213835768;T:255268464;N:59861 | 8 | 26 | 98 | 268758608 | 193670297 | 213835768 | 255268464 | 59861 | SRX21223187 | SRS18479990 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.9283 | 0.09341 | 0.82211 | 0.49121 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24712 | 24712 | SRR25492015 | SRX21223186 | SRS18479989 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S16 | GSM7676111 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S16 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676111 | GSM7676111: KBTGR cut replicate S16; Danio rerio; RNA Seq | GSM7676111 r1 | GSM7676111 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTGRcut_S16_L001_I1_001.fastq.gz H27K5BCX2_KBTGRcut_S16_L001_R1_001.fastq.gz H27K5BCX2_KBTGRcut_S16_L001_R2_001.fastq.gz | fastq fastq fastq | 1733777364.0 | 13134677.0 | GSM7676111 r1 | 0:8 1:26 2:98 | A:377310857;C:268033707;G:292815269;T:348639631;N:398882 | 8 | 26 | 98 | 377310857 | 268033707 | 292815269 | 348639631 | 398882 | SRX21223186 | SRS18479989 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92457 | 0.10972 | 0.83684 | 0.49054 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24713 | 24713 | SRR25492016 | SRX21223186 | SRS18479989 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S16 | GSM7676111 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S16 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676111 | GSM7676111: KBTGR cut replicate S16; Danio rerio; RNA Seq | GSM7676111 r1 | GSM7676111 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTGRcut_S16_L001_I1_001.fastq.gz HFJ5GBCX2_KBTGRcut_S16_L001_R1_001.fastq.gz HFJ5GBCX2_KBTGRcut_S16_L001_R2_001.fastq.gz | fastq fastq fastq | 1693931844.0 | 12832817.0 | GSM7676111 r2 | 0:8 1:26 2:98 | A:370335488;C:261641086;G:285793090;T:338853348;N:993054 | 8 | 26 | 98 | 370335488 | 261641086 | 285793090 | 338853348 | 993054 | SRX21223186 | SRS18479989 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.9257 | 0.11044 | 0.83593 | 0.49929 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24714 | 24714 | SRR25492017 | SRX21223186 | SRS18479989 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S16 | GSM7676111 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S16 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676111 | GSM7676111: KBTGR cut replicate S16; Danio rerio; RNA Seq | GSM7676111 r1 | GSM7676111 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTGRcut_S16_L001_I1_001.fastq.gz HYLGMBCXY_KBTGRcut_S16_L001_R1_001.fastq.gz HYLGMBCXY_KBTGRcut_S16_L001_R2_001.fastq.gz | fastq fastq fastq | 1822627488.0 | 13807784.0 | GSM7676111 r3 | 0:8 1:26 2:98 | A:396109320;C:281479505;G:307344748;T:362166256;N:6063003 | 8 | 26 | 98 | 396109320 | 281479505 | 307344748 | 362166256 | 6063003 | SRX21223186 | SRS18479989 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.9237 | 0.1075 | 0.83733 | 0.49995 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24715 | 24715 | SRR25492018 | SRX21223186 | SRS18479989 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S16 | GSM7676111 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S16 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676111 | GSM7676111: KBTGR cut replicate S16; Danio rerio; RNA Seq | GSM7676111 r1 | GSM7676111 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTGRcut_S16_L001_I1_001.fastq.gz HYWGVBCXY_KBTGRcut_S16_L001_R1_001.fastq.gz HYWGVBCXY_KBTGRcut_S16_L001_R2_001.fastq.gz | fastq fastq fastq | 1686941784.0 | 12779862.0 | GSM7676111 r4 | 0:8 1:26 2:98 | A:366995826;C:260455612;G:284984185;T:339737397;N:253456 | 8 | 26 | 98 | 366995826 | 260455612 | 284984185 | 339737397 | 253456 | SRX21223186 | SRS18479989 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92245 | 0.10934 | 0.83583 | 0.5109 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24716 | 24716 | SRR25492019 | SRX21223186 | SRS18479989 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S16 | GSM7676111 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S16 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676111 | GSM7676111: KBTGR cut replicate S16; Danio rerio; RNA Seq | GSM7676111 r1 | GSM7676111 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTGRcut_S16_L002_I1_001.fastq.gz HYLGMBCXY_KBTGRcut_S16_L002_R1_001.fastq.gz HYLGMBCXY_KBTGRcut_S16_L002_R2_001.fastq.gz | fastq fastq fastq | 1800605268.0 | 13640949.0 | GSM7676111 r7 | 0:8 1:26 2:98 | A:392033512;C:277884536;G:303332072;T:357657308;N:5905574 | 8 | 26 | 98 | 392033512 | 277884536 | 303332072 | 357657308 | 5905574 | SRX21223186 | SRS18479989 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92264 | 0.10865 | 0.83771 | 0.4962 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24717 | 24717 | SRR25492020 | SRX21223186 | SRS18479989 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S16 | GSM7676111 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S16 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676111 | GSM7676111: KBTGR cut replicate S16; Danio rerio; RNA Seq | GSM7676111 r1 | GSM7676111 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTGRcut_S16_L002_I1_001.fastq.gz HYWGVBCXY_KBTGRcut_S16_L002_R1_001.fastq.gz HYWGVBCXY_KBTGRcut_S16_L002_R2_001.fastq.gz | fastq fastq fastq | 1687212120.0 | 12781910.0 | GSM7676111 r8 | 0:8 1:26 2:98 | A:367591713;C:260374660;G:284790982;T:339790038;N:79787 | 8 | 26 | 98 | 367591713 | 260374660 | 284790982 | 339790038 | 79787 | SRX21223186 | SRS18479989 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92439 | 0.11008 | 0.83252 | 0.48566 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24718 | 24718 | SRR25492035 | SRX21223186 | SRS18479989 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S16 | GSM7676111 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S16 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676111 | GSM7676111: KBTGR cut replicate S16; Danio rerio; RNA Seq | GSM7676111 r1 | GSM7676111 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTGRcut_S16_L002_I1_001.fastq.gz H27K5BCX2_KBTGRcut_S16_L002_R1_001.fastq.gz H27K5BCX2_KBTGRcut_S16_L002_R2_001.fastq.gz | fastq fastq fastq | 1715599116.0 | 12996963.0 | GSM7676111 r5 | 0:8 1:26 2:98 | A:372113041;C:265506989;G:290255809;T:345508071;N:318464 | 8 | 26 | 98 | 372113041 | 265506989 | 290255809 | 345508071 | 318464 | SRX21223186 | SRS18479989 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92368 | 0.10681 | 0.83727 | 0.49306 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24719 | 24719 | SRR25492036 | SRX21223186 | SRS18479989 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S16 | GSM7676111 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S16 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676111 | GSM7676111: KBTGR cut replicate S16; Danio rerio; RNA Seq | GSM7676111 r1 | GSM7676111 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTGRcut_S16_L002_I1_001.fastq.gz HFJ5GBCX2_KBTGRcut_S16_L002_R1_001.fastq.gz HFJ5GBCX2_KBTGRcut_S16_L002_R2_001.fastq.gz | fastq fastq fastq | 1643667564.0 | 12452027.0 | GSM7676111 r6 | 0:8 1:26 2:98 | A:358646057;C:254091415;G:277620310;T:329013074;N:927790 | 8 | 26 | 98 | 358646057 | 254091415 | 277620310 | 329013074 | 927790 | SRX21223186 | SRS18479989 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92527 | 0.1079 | 0.83465 | 0.49184 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24720 | 24720 | SRR25492021 | SRX21223185 | SRS18479988 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE normal replicate S2 | GSM7676121 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE normal replicate S2 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676121 | GSM7676121: KBTRE normal replicate S2; Danio rerio; RNA Seq | GSM7676121 r1 | GSM7676121 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTREnormal_S2_L001_I1_001.fastq.gz H27K5BCX2_KBTREnormal_S2_L001_R1_001.fastq.gz H27K5BCX2_KBTREnormal_S2_L001_R2_001.fastq.gz | fastq fastq fastq | 807188844.0 | 6115067.0 | GSM7676121 r1 | 0:8 1:26 2:98 | A:172308620;C:125913345;G:139442237;T:161426336;N:186028 | 8 | 26 | 98 | 172308620 | 125913345 | 139442237 | 161426336 | 186028 | SRX21223185 | SRS18479988 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92298 | 0.08811 | 0.85433 | 0.4736 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24721 | 24721 | SRR25492022 | SRX21223185 | SRS18479988 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE normal replicate S2 | GSM7676121 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE normal replicate S2 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676121 | GSM7676121: KBTRE normal replicate S2; Danio rerio; RNA Seq | GSM7676121 r1 | GSM7676121 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTREnormal_S2_L001_I1_001.fastq.gz HFJ5GBCX2_KBTREnormal_S2_L001_R1_001.fastq.gz HFJ5GBCX2_KBTREnormal_S2_L001_R2_001.fastq.gz | fastq fastq fastq | 793147344.0 | 6008692.0 | GSM7676121 r2 | 0:8 1:26 2:98 | A:169497439;C:123759266;G:137078289;T:158052236;N:464586 | 8 | 26 | 98 | 169497439 | 123759266 | 137078289 | 158052236 | 464586 | SRX21223185 | SRS18479988 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92361 | 0.08644 | 0.85464 | 0.47495 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24722 | 24722 | SRR25492023 | SRX21223185 | SRS18479988 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE normal replicate S2 | GSM7676121 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE normal replicate S2 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676121 | GSM7676121: KBTRE normal replicate S2; Danio rerio; RNA Seq | GSM7676121 r1 | GSM7676121 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTREnormal_S2_L001_I1_001.fastq.gz HYLGMBCXY_KBTREnormal_S2_L001_R1_001.fastq.gz HYLGMBCXY_KBTREnormal_S2_L001_R2_001.fastq.gz | fastq fastq fastq | 852294828.0 | 6456779.0 | GSM7676121 r3 | 0:8 1:26 2:98 | A:181361509;C:132917413;G:147157966;T:168488477;N:2838977 | 8 | 26 | 98 | 181361509 | 132917413 | 147157966 | 168488477 | 2838977 | SRX21223185 | SRS18479988 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.9234 | 0.08441 | 0.85681 | 0.45235 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24723 | 24723 | SRR25492024 | SRX21223185 | SRS18479988 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE normal replicate S2 | GSM7676121 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE normal replicate S2 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676121 | GSM7676121: KBTRE normal replicate S2; Danio rerio; RNA Seq | GSM7676121 r1 | GSM7676121 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTREnormal_S2_L001_I1_001.fastq.gz HYWGVBCXY_KBTREnormal_S2_L001_R1_001.fastq.gz HYWGVBCXY_KBTREnormal_S2_L001_R2_001.fastq.gz | fastq fastq fastq | 781758516.0 | 5922413.0 | GSM7676121 r4 | 0:8 1:26 2:98 | A:166724753;C:121839518;G:135085811;T:156626231;N:120161 | 8 | 26 | 98 | 166724753 | 121839518 | 135085811 | 156626231 | 120161 | SRX21223185 | SRS18479988 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92293 | 0.08698 | 0.85626 | 0.47803 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24724 | 24724 | SRR25492025 | SRX21223185 | SRS18479988 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE normal replicate S2 | GSM7676121 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE normal replicate S2 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676121 | GSM7676121: KBTRE normal replicate S2; Danio rerio; RNA Seq | GSM7676121 r1 | GSM7676121 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTREnormal_S2_L002_I1_001.fastq.gz HYLGMBCXY_KBTREnormal_S2_L002_R1_001.fastq.gz HYLGMBCXY_KBTREnormal_S2_L002_R2_001.fastq.gz | fastq fastq fastq | 843137856.0 | 6387408.0 | GSM7676121 r7 | 0:8 1:26 2:98 | A:179558862;C:131385546;G:145540868;T:166696387;N:2784321 | 8 | 26 | 98 | 179558862 | 131385546 | 145540868 | 166696387 | 2784321 | SRX21223185 | SRS18479988 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92281 | 0.08446 | 0.85685 | 0.47524 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24725 | 24725 | SRR25492026 | SRX21223185 | SRS18479988 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE normal replicate S2 | GSM7676121 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE normal replicate S2 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676121 | GSM7676121: KBTRE normal replicate S2; Danio rerio; RNA Seq | GSM7676121 r1 | GSM7676121 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTREnormal_S2_L002_I1_001.fastq.gz HYWGVBCXY_KBTREnormal_S2_L002_R1_001.fastq.gz HYWGVBCXY_KBTREnormal_S2_L002_R2_001.fastq.gz | fastq fastq fastq | 782789964.0 | 5930227.0 | GSM7676121 r8 | 0:8 1:26 2:98 | A:167092677;C:121959994;G:135189997;T:156882004;N:37574 | 8 | 26 | 98 | 167092677 | 121959994 | 135189997 | 156882004 | 37574 | SRX21223185 | SRS18479988 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92384 | 0.08593 | 0.85529 | 0.47658 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24726 | 24726 | SRR25492073 | SRX21223185 | SRS18479988 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE normal replicate S2 | GSM7676121 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE normal replicate S2 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676121 | GSM7676121: KBTRE normal replicate S2; Danio rerio; RNA Seq | GSM7676121 r1 | GSM7676121 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTREnormal_S2_L002_I1_001.fastq.gz H27K5BCX2_KBTREnormal_S2_L002_R1_001.fastq.gz H27K5BCX2_KBTREnormal_S2_L002_R2_001.fastq.gz | fastq fastq fastq | 799492452.0 | 6056761.0 | GSM7676121 r5 | 0:8 1:26 2:98 | A:170481537;C:124727025;G:138222223;T:159980825;N:150968 | 8 | 26 | 98 | 170481537 | 124727025 | 138222223 | 159980825 | 150968 | SRX21223185 | SRS18479988 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92219 | 0.08693 | 0.85689 | 0.45239 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24727 | 24727 | SRR25492074 | SRX21223185 | SRS18479988 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTRE normal replicate S2 | GSM7676121 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTRE normal replicate S2 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676121 | GSM7676121: KBTRE normal replicate S2; Danio rerio; RNA Seq | GSM7676121 r1 | GSM7676121 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTREnormal_S2_L002_I1_001.fastq.gz HFJ5GBCX2_KBTREnormal_S2_L002_R1_001.fastq.gz HFJ5GBCX2_KBTREnormal_S2_L002_R2_001.fastq.gz | fastq fastq fastq | 771619332.0 | 5845601.0 | GSM7676121 r6 | 0:8 1:26 2:98 | A:164827141;C:120458327;G:133444180;T:153706468;N:432782 | 8 | 26 | 98 | 164827141 | 120458327 | 133444180 | 153706468 | 432782 | SRX21223185 | SRS18479988 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.9244 | 0.086 | 0.85752 | 0.46487 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24728 | 24728 | SRR25492027 | SRX21223184 | SRS18479986 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S15 | GSM7676110 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S15 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676110 | GSM7676110: KBTGR cut replicate S15; Danio rerio; RNA Seq | GSM7676110 r1 | GSM7676110 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTGRcut_S15_L001_I1_001.fastq.gz H27K5BCX2_KBTGRcut_S15_L001_R1_001.fastq.gz H27K5BCX2_KBTGRcut_S15_L001_R2_001.fastq.gz | fastq fastq fastq | 991634028.0 | 7512379.0 | GSM7676110 r1 | 0:8 1:26 2:98 | A:213910719;C:153374311;G:167803904;T:200905324;N:218884 | 8 | 26 | 98 | 213910719 | 153374311 | 167803904 | 200905324 | 218884 | SRX21223184 | SRS18479986 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92289 | 0.10915 | 0.83163 | 0.51061 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24729 | 24729 | SRR25492028 | SRX21223184 | SRS18479986 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S15 | GSM7676110 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S15 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676110 | GSM7676110: KBTGR cut replicate S15; Danio rerio; RNA Seq | GSM7676110 r1 | GSM7676110 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTGRcut_S15_L001_I1_001.fastq.gz HFJ5GBCX2_KBTGRcut_S15_L001_R1_001.fastq.gz HFJ5GBCX2_KBTGRcut_S15_L001_R2_001.fastq.gz | fastq fastq fastq | 973359420.0 | 7373935.0 | GSM7676110 r2 | 0:8 1:26 2:98 | A:210076453;C:150636615;G:164768683;T:196577221;N:586658 | 8 | 26 | 98 | 210076453 | 150636615 | 164768683 | 196577221 | 586658 | SRX21223184 | SRS18479986 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92307 | 0.10829 | 0.8309 | 0.50664 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24730 | 24730 | SRR25492029 | SRX21223184 | SRS18479986 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S15 | GSM7676110 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S15 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676110 | GSM7676110: KBTGR cut replicate S15; Danio rerio; RNA Seq | GSM7676110 r1 | GSM7676110 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTGRcut_S15_L001_I1_001.fastq.gz HYLGMBCXY_KBTGRcut_S15_L001_R1_001.fastq.gz HYLGMBCXY_KBTGRcut_S15_L001_R2_001.fastq.gz | fastq fastq fastq | 1041506532.0 | 7890201.0 | GSM7676110 r3 | 0:8 1:26 2:98 | A:224331472;C:160956885;G:176036393;T:208465176;N:3449772 | 8 | 26 | 98 | 224331472 | 160956885 | 176036393 | 208465176 | 3449772 | SRX21223184 | SRS18479986 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92046 | 0.10509 | 0.83179 | 0.5158 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24731 | 24731 | SRR25492030 | SRX21223184 | SRS18479986 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S15 | GSM7676110 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S15 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676110 | GSM7676110: KBTGR cut replicate S15; Danio rerio; RNA Seq | GSM7676110 r1 | GSM7676110 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTGRcut_S15_L001_I1_001.fastq.gz HYWGVBCXY_KBTGRcut_S15_L001_R1_001.fastq.gz HYWGVBCXY_KBTGRcut_S15_L001_R2_001.fastq.gz | fastq fastq fastq | 959532816.0 | 7269188.0 | GSM7676110 r4 | 0:8 1:26 2:98 | A:207101192;C:148189510;G:162346748;T:194597135;N:145839 | 8 | 26 | 98 | 207101192 | 148189510 | 162346748 | 194597135 | 145839 | SRX21223184 | SRS18479986 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92159 | 0.10724 | 0.83031 | 0.51032 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24732 | 24732 | SRR25492031 | SRX21223184 | SRS18479986 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S15 | GSM7676110 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S15 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676110 | GSM7676110: KBTGR cut replicate S15; Danio rerio; RNA Seq | GSM7676110 r1 | GSM7676110 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTGRcut_S15_L002_I1_001.fastq.gz H27K5BCX2_KBTGRcut_S15_L002_R1_001.fastq.gz H27K5BCX2_KBTGRcut_S15_L002_R2_001.fastq.gz | fastq fastq fastq | 981662748.0 | 7436839.0 | GSM7676110 r5 | 0:8 1:26 2:98 | A:211518289;C:151920644;G:166249815;T:198931555;N:189919 | 8 | 26 | 98 | 211518289 | 151920644 | 166249815 | 198931555 | 189919 | SRX21223184 | SRS18479986 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92149 | 0.10822 | 0.83078 | 0.51544 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24733 | 24733 | SRR25492032 | SRX21223184 | SRS18479986 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S15 | GSM7676110 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S15 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676110 | GSM7676110: KBTGR cut replicate S15; Danio rerio; RNA Seq | GSM7676110 r1 | GSM7676110 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTGRcut_S15_L002_I1_001.fastq.gz HFJ5GBCX2_KBTGRcut_S15_L002_R1_001.fastq.gz HFJ5GBCX2_KBTGRcut_S15_L002_R2_001.fastq.gz | fastq fastq fastq | 945758088.0 | 7164834.0 | GSM7676110 r6 | 0:8 1:26 2:98 | A:203967307;C:146422295;G:160254376;T:190978208;N:531546 | 8 | 26 | 98 | 203967307 | 146422295 | 160254376 | 190978208 | 531546 | SRX21223184 | SRS18479986 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92245 | 0.10759 | 0.83142 | 0.51632 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24734 | 24734 | SRR25492033 | SRX21223184 | SRS18479986 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S15 | GSM7676110 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S15 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676110 | GSM7676110: KBTGR cut replicate S15; Danio rerio; RNA Seq | GSM7676110 r1 | GSM7676110 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTGRcut_S15_L002_I1_001.fastq.gz HYLGMBCXY_KBTGRcut_S15_L002_R1_001.fastq.gz HYLGMBCXY_KBTGRcut_S15_L002_R2_001.fastq.gz | fastq fastq fastq | 1029633660.0 | 7800255.0 | GSM7676110 r7 | 0:8 1:26 2:98 | A:221920698;C:159045837;G:173908417;T:206140930;N:3409108 | 8 | 26 | 98 | 221920698 | 159045837 | 173908417 | 206140930 | 3409108 | SRX21223184 | SRS18479986 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92088 | 0.10596 | 0.83295 | 0.51602 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24735 | 24735 | SRR25492034 | SRX21223184 | SRS18479986 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S15 | GSM7676110 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S15 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676110 | GSM7676110: KBTGR cut replicate S15; Danio rerio; RNA Seq | GSM7676110 r1 | GSM7676110 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTGRcut_S15_L002_I1_001.fastq.gz HYWGVBCXY_KBTGRcut_S15_L002_R1_001.fastq.gz HYWGVBCXY_KBTGRcut_S15_L002_R2_001.fastq.gz | fastq fastq fastq | 960845292.0 | 7279131.0 | GSM7676110 r8 | 0:8 1:26 2:98 | A:207492886;C:148360000;G:162496060;T:194960451;N:45441 | 8 | 26 | 98 | 207492886 | 148360000 | 162496060 | 194960451 | 45441 | SRX21223184 | SRS18479986 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92106 | 0.10819 | 0.82946 | 0.51364 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24736 | 24736 | SRR25492037 | SRX21223183 | SRS18479987 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S14 | GSM7676109 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S14 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676109 | GSM7676109: KBTGR cut replicate S14; Danio rerio; RNA Seq | GSM7676109 r1 | GSM7676109 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTGRcut_S14_L001_I1_001.fastq.gz H27K5BCX2_KBTGRcut_S14_L001_R1_001.fastq.gz H27K5BCX2_KBTGRcut_S14_L001_R2_001.fastq.gz | fastq fastq fastq | 2591037900.0 | 19629075.0 | GSM7676109 r1 | 0:8 1:26 2:98 | A:555580990;C:402990210;G:441965039;T:522520043;N:593068 | 8 | 26 | 98 | 555580990 | 402990210 | 441965039 | 522520043 | 593068 | SRX21223183 | SRS18479987 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92241 | 0.10605 | 0.83187 | 0.51181 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24737 | 24737 | SRR25492038 | SRX21223183 | SRS18479987 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S14 | GSM7676109 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S14 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676109 | GSM7676109: KBTGR cut replicate S14; Danio rerio; RNA Seq | GSM7676109 r1 | GSM7676109 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTGRcut_S14_L001_I1_001.fastq.gz HFJ5GBCX2_KBTGRcut_S14_L001_R1_001.fastq.gz HFJ5GBCX2_KBTGRcut_S14_L001_R2_001.fastq.gz | fastq fastq fastq | 2497809996.0 | 18922803.0 | GSM7676109 r2 | 0:8 1:26 2:98 | A:535857014;C:388641161;G:426148721;T:502295548;N:1492250 | 8 | 26 | 98 | 535857014 | 388641161 | 426148721 | 502295548 | 1492250 | SRX21223183 | SRS18479987 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92376 | 0.10611 | 0.83177 | 0.49598 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24738 | 24738 | SRR25492039 | SRX21223183 | SRS18479987 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S14 | GSM7676109 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S14 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676109 | GSM7676109: KBTGR cut replicate S14; Danio rerio; RNA Seq | GSM7676109 r1 | GSM7676109 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTGRcut_S14_L001_I1_001.fastq.gz HYLGMBCXY_KBTGRcut_S14_L001_R1_001.fastq.gz HYLGMBCXY_KBTGRcut_S14_L001_R2_001.fastq.gz | fastq fastq fastq | 2728654500.0 | 20671625.0 | GSM7676109 r3 | 0:8 1:26 2:98 | A:584168882;C:423983386;G:464567203;T:543992364;N:9107415 | 8 | 26 | 98 | 584168882 | 423983386 | 464567203 | 543992364 | 9107415 | SRX21223183 | SRS18479987 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92388 | 0.10372 | 0.83181 | 0.50245 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24739 | 24739 | SRR25492040 | SRX21223183 | SRS18479987 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S14 | GSM7676109 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S14 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676109 | GSM7676109: KBTGR cut replicate S14; Danio rerio; RNA Seq | GSM7676109 r1 | GSM7676109 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTGRcut_S14_L001_I1_001.fastq.gz HYWGVBCXY_KBTGRcut_S14_L001_R1_001.fastq.gz HYWGVBCXY_KBTGRcut_S14_L001_R2_001.fastq.gz | fastq fastq fastq | 2618092488.0 | 19834034.0 | GSM7676109 r4 | 0:8 1:26 2:98 | A:561803683;C:406792207;G:446399294;T:528347486;N:392662 | 8 | 26 | 98 | 561803683 | 406792207 | 446399294 | 528347486 | 392662 | SRX21223183 | SRS18479987 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92126 | 0.10547 | 0.83161 | 0.49641 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24740 | 24740 | SRR25492041 | SRX21223183 | SRS18479987 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S14 | GSM7676109 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S14 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676109 | GSM7676109: KBTGR cut replicate S14; Danio rerio; RNA Seq | GSM7676109 r1 | GSM7676109 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | H27K5BCX2_KBTGRcut_S14_L002_I1_001.fastq.gz H27K5BCX2_KBTGRcut_S14_L002_R1_001.fastq.gz H27K5BCX2_KBTGRcut_S14_L002_R2_001.fastq.gz | fastq fastq fastq | 2564163096.0 | 19425478.0 | GSM7676109 r5 | 0:8 1:26 2:98 | A:549889204;C:398857562;G:437417974;T:517052381;N:479723 | 8 | 26 | 98 | 549889204 | 398857562 | 437417974 | 517052381 | 479723 | SRX21223183 | SRS18479987 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.9232 | 0.1061 | 0.83071 | 0.50282 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24741 | 24741 | SRR25492042 | SRX21223183 | SRS18479987 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S14 | GSM7676109 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S14 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676109 | GSM7676109: KBTGR cut replicate S14; Danio rerio; RNA Seq | GSM7676109 r1 | GSM7676109 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HFJ5GBCX2_KBTGRcut_S14_L002_I1_001.fastq.gz HFJ5GBCX2_KBTGRcut_S14_L002_R1_001.fastq.gz HFJ5GBCX2_KBTGRcut_S14_L002_R2_001.fastq.gz | fastq fastq fastq | 2415052992.0 | 18295856.0 | GSM7676109 r6 | 0:8 1:26 2:98 | A:517947164;C:375865420;G:412233894;T:485594270;N:1353140 | 8 | 26 | 98 | 517947164 | 375865420 | 412233894 | 485594270 | 1353140 | SRX21223183 | SRS18479987 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92288 | 0.10488 | 0.82899 | 0.4957 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24742 | 24742 | SRR25492043 | SRX21223183 | SRS18479987 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S14 | GSM7676109 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S14 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676109 | GSM7676109: KBTGR cut replicate S14; Danio rerio; RNA Seq | GSM7676109 r1 | GSM7676109 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYLGMBCXY_KBTGRcut_S14_L002_I1_001.fastq.gz HYLGMBCXY_KBTGRcut_S14_L002_R1_001.fastq.gz HYLGMBCXY_KBTGRcut_S14_L002_R2_001.fastq.gz | fastq fastq fastq | 2694434160.0 | 20412380.0 | GSM7676109 r7 | 0:8 1:26 2:98 | A:576891044;C:418621857;G:458706495;T:537348616;N:8845228 | 8 | 26 | 98 | 576891044 | 418621857 | 458706495 | 537348616 | 8845228 | SRX21223183 | SRS18479987 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.9222 | 0.10385 | 0.83114 | 0.50871 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures | |||||||||||||||||
| 24743 | 24743 | SRR25492044 | SRX21223183 | SRS18479987 | SRP452960 | PRJNA1001330 | Ontogenetically distinct neutrophils differ in function and transcriptional profile in zebrafish | GSE239880 | Transcriptome Analysis | The current view of hematopoiesis considers leukocytes on a continuum with distinct developmental origins and which exert non overlapping functions. However there is little information about ontogenetically distinct neutrophil populations. In this work using a photoconvertible transgenic zebrafish line; Tgmpx:Dendra2 we selectively label rostral blood island derived and caudal hematopoietic tissue derived neutrophils in vivo during steady state or upon injury. By comparing the migratory properties and single cell expression profiles of both neutrophil populations at steady state we reveal that rostral neutrophils show higher csf3b expression and migration capacity than caudal neutrophils. Upon injury both populations share a core transcriptional profile as well as subset specific transcriptional signatures. Accordingly both rostral and caudal neutrophils are recruited to the wound independently of their distance to the injury. While rostral neutrophils respond uniformly caudal neutrophils respond heterogeneously. Collectively our results reveal that co existing neutrophils populations with ontogenically distinct origin display key functional differences. Overall design: We photoconverted RBI derived neutrophils in the head two times at 34hpf and 44hpf and performed caudal fin transections at 54hpf on the half of each group. Three hours later we homogenized non cut and cut embryos from each group and separately sorted red and green fluorescent cells. The resulting four groups were analized by scRNAseq. | KBTGR cut replicate S14 | GSM7676109 | source name:whole embryo|tissue:whole embryo|cell type:neutrophils|geo loc name:missing|collection date:missing | KBTGR cut replicate S14 | Reads from the four separate conditions flowcells were preprocessed using Seurat in R https://satijalab.org/seurat/ Assembly: GRCz11 Supplementary files format and content: tab separated values files | whole embryo | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell 3’ Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | tissue:whole embryo|cell type:neutrophils | GSM7676109 | GSM7676109: KBTGR cut replicate S14; Danio rerio; RNA Seq | GSM7676109 r1 | GSM7676109 | 1 | For tissue disaggregation whole embryos were incubated with a digestion cocktail 10mg/ml collagenase; 5mg/ml hyaluronidase; and 20mg/ml proteinase K and constant mechanical dissociation was exerted for 10 minutes. Cells suspension were filtered through a 40 µm nylon mesh and L 15 medium was added to stop the digestion process. Isolated cells were pelleted resuspended in L 15 medium and processed by FACS. Red+ and green+ cells were loaded into the Chromium Controller instrument to generate Gel beads in EMulsion GEMs. GEM RT was performed in a C1000 Touch Thermal cycler. post reverse transcription GEMs were broken and the single strand cDNA was cleaned up using DynaBeads MyOne Silane Beads and the SPRIselect Reagent Kit. cDNA was amplified using the C1000 Touch Thermal cycler and cleaned up with the SPRIselect Reagent Kit. Libraries were constructed using the GemCode Single Cell three prime Library Kit following these steps: 1 end repair and A tailing 2 adapter ligation 3 postligation cleanup with SPRIselect 4 sample index PCR and cleanup. The barcode sequencing libraries were quantified by quantitative PCR. | RNA-Seq | TRANSCRIPTOMIC SINGLE CELL | cDNA | PAIRED | ILLUMINA | Illumina HiSeq 4000 | SRP452960 | loader:fastq load.py | HYWGVBCXY_KBTGRcut_S14_L002_I1_001.fastq.gz HYWGVBCXY_KBTGRcut_S14_L002_R1_001.fastq.gz HYWGVBCXY_KBTGRcut_S14_L002_R2_001.fastq.gz | fastq fastq fastq | 2615928216.0 | 19817638.0 | GSM7676109 r8 | 0:8 1:26 2:98 | A:561516458;C:406357695;G:445851758;T:528279876;N:122737 | 8 | 26 | 98 | 561516458 | 406357695 | 445851758 | 528279876 | 122737 | SRX21223183 | SRS18479987 | SRA1685258 | Facultad de Ciencias de la vida, Universidad Andres Bello | Facultad de Ciencias de la vida, Universidad Andres Bello | 1 | 0.92228 | 0.10465 | 0.8297 | 0.49948 | 98 | B | usable mapping rate | illumina | hiseq_era | unknown | random_priming | unknown | sc | single_cell_droplet | 10x | Chile | 2023-08-02 | Undetermined | Embryo | Whole Organism | All anatomical structures |
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CREATE TABLE run_metadata("run.accession" VARCHAR, "experiment.accession" VARCHAR, "sample.accession" VARCHAR, "study.accession" VARCHAR, bioproject VARCHAR, "study.title" VARCHAR, "study.alias" VARCHAR, "study.type" VARCHAR, "study.abstract" VARCHAR, "study.attributes" VARCHAR, "study.PMIDs" VARCHAR, "sample.description" VARCHAR, "sample.title" VARCHAR, "sample.alias" VARCHAR, "sample.centername" VARCHAR, "sample.attributes" VARCHAR, "GEOsample.title" VARCHAR, "GEOsample.dataprocessing" VARCHAR, "GEOsample.source" VARCHAR, "GEOsample.treatmentprotocol" VARCHAR, "GEOsample.extractprotocol" VARCHAR, "GEOsample.growthprotocol" VARCHAR, "GEOsample.characteristics" VARCHAR, "GEOsample.accession" VARCHAR, "experiment.title" VARCHAR, "experiment.alias" VARCHAR, "experiment.library_name" VARCHAR, "experiment.design_description" VARCHAR, "experiment.library_construction_protocol" VARCHAR, "experiment.attributes" VARCHAR, "experiment.library_strategy" VARCHAR, "experiment.library_source" VARCHAR, "experiment.library_selection" VARCHAR, "experiment.library_layout" VARCHAR, "experiment.platform" VARCHAR, "experiment.instrument_model" VARCHAR, "experiment.spot_descriptor" VARCHAR, "experiment.study_ref" VARCHAR, "run.title" VARCHAR, "run.attributes" VARCHAR, "run.filename" VARCHAR, "run.semantic_name" VARCHAR, "run.total_bases" DOUBLE, "run.total_spots" DOUBLE, "run.alias" VARCHAR, "run.read_lengths" VARCHAR, "run.base_counts" VARCHAR, "run.r1_length" BIGINT, "run.r2_length" BIGINT, "run.r3_length" BIGINT, "run.r4_length" BIGINT, "run.Acount" BIGINT, "run.Ccount" BIGINT, "run.Gcount" BIGINT, "run.Tcount" BIGINT, "run.Ncount" BIGINT, "run.experiment" VARCHAR, "run.pool_member" VARCHAR, "submission.accession" VARCHAR, "submission.srasource" VARCHAR, "submission.bioprojectsource" VARCHAR, "seqdetective.n_mates" BIGINT, "seqdetective.mapping_rate.mate1" DOUBLE, "seqdetective.mapping_rate.mate2" DOUBLE, "seqdetective.nofeature_rate.mate1" DOUBLE, "seqdetective.nofeature_rate.mate2" DOUBLE, "seqdetective.sparsity.mate1" DOUBLE, "seqdetective.sparsity.mate2" DOUBLE, "seqdetective.pos_strand_rate.mate1" DOUBLE, "seqdetective.pos_strand_rate.mate2" DOUBLE, "seqdetective.readlen.mate1" BIGINT, "seqdetective.readlen.mate2" BIGINT, "seqdetective.judgement.mate1" VARCHAR, "seqdetective.judgement.mate2" VARCHAR, "seqdetective.judgement.reason" VARCHAR, platform_family VARCHAR, instrument_generation VARCHAR, read_bias VARCHAR, selection_class VARCHAR, prep_kit VARCHAR, sc_or_bulk VARCHAR, tech_class VARCHAR, technology VARCHAR, tech_variant VARCHAR, "submission.bioprojectsource.country" VARCHAR, earliest_date DATE, devstage_curation VARCHAR, devstage_curation_coarse VARCHAR, tissue_curation VARCHAR, tissue_curation_coarse VARCHAR);;
CREATE INDEX idx_run_bioproject ON run_metadata(bioproject);;
CREATE INDEX idx_run_run_accession ON run_metadata("run.accession");;