{"database": "metadata", "table": "run_metadata", "is_view": false, "human_description_en": "where experiment.library_layout = \"SINGLE\", experiment.library_strategy = \"RNA-Seq\" and technology = \"celseq\"", "rows": [[10060, "ERR4795364", "ERX4665135", "ERS5281176", "ERP124847", "PRJEB41113", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E-MTAB-9727", "Transcriptome Analysis", "The mesothelium forms epithelial membranes that line the bodies cavities and surround the internal organs. Mesothelia widely contribute to organ homeostasis and regeneration  and their dysregulation can result in congenital anomalies of the viscera  ventral wall defects  and mesothelioma tumors. Nonetheless  the embryonic ontogeny and developmental regulation of mesothelium formation has remained uncharted. Here  we combine genetic lineage tracing  in toto live imaging  and single cell transcriptomics in zebrafish to track mesothelial progenitor origins from the lateral plate mesoderm LPM. Our single cell analysis uncovers a post gastrulation gene expression signature centered on hand2 that delineates distinct progenitor populations within the forming LPM. Combining gene expression analysis and imaging of transgenic reporter zebrafish embryos  we chart the origin of mesothelial progenitors to the lateral most  hand2 expressing LPM and confirm evolutionary conservation in mouse. Our time lapse imaging of transgenic hand2 reporter embryos captures zebrafish mesothelium formation  documenting the coordinated cell movements that form pericardium and visceral and parietal peritoneum. We establish that the primordial germ cells migrate associated with the forming mesothelium as ventral migration boundary. Functionally  hand2 mutants fail to close the ventral mesothelium due to perturbed migration of mesothelium progenitors. Analyzing mouse and human mesothelioma tumors hypothesized to emerge from transformed mesothelium  we find de novo expression of LPM associated transcription factors  and in particular of Hand2  indicating the re initiation of a developmental transcriptional program in mesothelioma. Taken together  our work outlines a genetic and developmental signature of mesothelial origins centered around Hand2  contributing to our understanding of mesothelial pathologies and mesothelioma.", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", null, "Protocols: Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Sample 4", "E MTAB 9727:Sample 4", null, "isolate:not applicable|organism:Danio rerio|sex:mixed|age:10|developmental stage:tailbud stage|organism part:lateral plate mesoderm plate|genotype:drl:mCherry|ENA FIRST PUBLIC:2021 04 01|ENA LAST UPDATE:2021 09 01", null, null, null, null, null, null, null, null, "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E MTAB 9727:Sample 4 s", "Sample 4 s", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Experimental Factor: replicate:lateral plate mesoderm plate 4", "RNA-Seq", "TRANSCRIPTOMIC SINGLE CELL", "RANDOM", "SINGLE", "ILLUMINA", "NextSeq 500", null, "ERP124847", "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", "SN4_AH2TW3BGX5_S1_R1_cat.fastq.gz", "fastq", 5410723202.0, 71707309.0, "E MTAB 9727:Sample 4", "0:75.46 1:0", "A:1330416803;C:531269504;G:734031980;T:2814959249;N:45666", 75, 0, null, null, 1330416803, 531269504, 734031980, 2814959249, 45666, "ERX4665135", "ERS5281176", "ERA3048543", "University of Zurich|European Nucleotide Archive", "University of Zurich|European Nucleotide Archive", 1, 0.33614, null, 0.21532, null, 0.99019, null, 0.41002, null, 76, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Switzerland", "2021-04-01", "Gastrula", "Embryo", "Embryo Imprecise", "All anatomical structures"], [10061, "ERR4795365", "ERX4665135", "ERS5281176", "ERP124847", "PRJEB41113", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E-MTAB-9727", "Transcriptome Analysis", "The mesothelium forms epithelial membranes that line the bodies cavities and surround the internal organs. Mesothelia widely contribute to organ homeostasis and regeneration  and their dysregulation can result in congenital anomalies of the viscera  ventral wall defects  and mesothelioma tumors. Nonetheless  the embryonic ontogeny and developmental regulation of mesothelium formation has remained uncharted. Here  we combine genetic lineage tracing  in toto live imaging  and single cell transcriptomics in zebrafish to track mesothelial progenitor origins from the lateral plate mesoderm LPM. Our single cell analysis uncovers a post gastrulation gene expression signature centered on hand2 that delineates distinct progenitor populations within the forming LPM. Combining gene expression analysis and imaging of transgenic reporter zebrafish embryos  we chart the origin of mesothelial progenitors to the lateral most  hand2 expressing LPM and confirm evolutionary conservation in mouse. Our time lapse imaging of transgenic hand2 reporter embryos captures zebrafish mesothelium formation  documenting the coordinated cell movements that form pericardium and visceral and parietal peritoneum. We establish that the primordial germ cells migrate associated with the forming mesothelium as ventral migration boundary. Functionally  hand2 mutants fail to close the ventral mesothelium due to perturbed migration of mesothelium progenitors. Analyzing mouse and human mesothelioma tumors hypothesized to emerge from transformed mesothelium  we find de novo expression of LPM associated transcription factors  and in particular of Hand2  indicating the re initiation of a developmental transcriptional program in mesothelioma. Taken together  our work outlines a genetic and developmental signature of mesothelial origins centered around Hand2  contributing to our understanding of mesothelial pathologies and mesothelioma.", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", null, "Protocols: Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Sample 4", "E MTAB 9727:Sample 4", null, "isolate:not applicable|organism:Danio rerio|sex:mixed|age:10|developmental stage:tailbud stage|organism part:lateral plate mesoderm plate|genotype:drl:mCherry|ENA FIRST PUBLIC:2021 04 01|ENA LAST UPDATE:2021 09 01", null, null, null, null, null, null, null, null, "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E MTAB 9727:Sample 4 s", "Sample 4 s", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Experimental Factor: replicate:lateral plate mesoderm plate 4", "RNA-Seq", "TRANSCRIPTOMIC SINGLE CELL", "RANDOM", "SINGLE", "ILLUMINA", "NextSeq 500", null, "ERP124847", "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", "SN4_AH2TW3BGX5_S1_R2_cat.fastq.gz", "fastq", 5414184547.0, 71707309.0, "E MTAB 9727:Sample 4 1", "0:0 1:75.50", "A:1582938104;C:1052565419;G:1177180395;T:1600161662;N:1338967", 0, 75, null, null, 1582938104, 1052565419, 1177180395, 1600161662, 1338967, "ERX4665135", "ERS5281176", "ERA3048543", "University of Zurich|European Nucleotide Archive", "University of Zurich|European Nucleotide Archive", 1, 0.84976, null, 0.28521, null, 0.85038, null, 0.5124, null, 75, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Switzerland", "2021-04-01", "Gastrula", "Embryo", "Embryo Imprecise", "All anatomical structures"], [10062, "ERR4795362", "ERX4665134", "ERS5281175", "ERP124847", "PRJEB41113", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E-MTAB-9727", "Transcriptome Analysis", "The mesothelium forms epithelial membranes that line the bodies cavities and surround the internal organs. Mesothelia widely contribute to organ homeostasis and regeneration  and their dysregulation can result in congenital anomalies of the viscera  ventral wall defects  and mesothelioma tumors. Nonetheless  the embryonic ontogeny and developmental regulation of mesothelium formation has remained uncharted. Here  we combine genetic lineage tracing  in toto live imaging  and single cell transcriptomics in zebrafish to track mesothelial progenitor origins from the lateral plate mesoderm LPM. Our single cell analysis uncovers a post gastrulation gene expression signature centered on hand2 that delineates distinct progenitor populations within the forming LPM. Combining gene expression analysis and imaging of transgenic reporter zebrafish embryos  we chart the origin of mesothelial progenitors to the lateral most  hand2 expressing LPM and confirm evolutionary conservation in mouse. Our time lapse imaging of transgenic hand2 reporter embryos captures zebrafish mesothelium formation  documenting the coordinated cell movements that form pericardium and visceral and parietal peritoneum. We establish that the primordial germ cells migrate associated with the forming mesothelium as ventral migration boundary. Functionally  hand2 mutants fail to close the ventral mesothelium due to perturbed migration of mesothelium progenitors. Analyzing mouse and human mesothelioma tumors hypothesized to emerge from transformed mesothelium  we find de novo expression of LPM associated transcription factors  and in particular of Hand2  indicating the re initiation of a developmental transcriptional program in mesothelioma. Taken together  our work outlines a genetic and developmental signature of mesothelial origins centered around Hand2  contributing to our understanding of mesothelial pathologies and mesothelioma.", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", null, "Protocols: Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Sample 3", "E MTAB 9727:Sample 3", null, "isolate:not applicable|organism:Danio rerio|sex:mixed|age:10|developmental stage:tailbud stage|organism part:lateral plate mesoderm plate|genotype:drl:mCherry|ENA FIRST PUBLIC:2021 04 01|ENA LAST UPDATE:2021 09 01", null, null, null, null, null, null, null, null, "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E MTAB 9727:Sample 3 s", "Sample 3 s", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Experimental Factor: replicate:lateral plate mesoderm plate 3", "RNA-Seq", "TRANSCRIPTOMIC SINGLE CELL", "RANDOM", "SINGLE", "ILLUMINA", "NextSeq 500", null, "ERP124847", "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", "SN3_AHY3WGBGX3_S7_R1_cat.fastq.gz", "fastq", 4823193891.0, 63965519.0, "E MTAB 9727:Sample 3", "0:75.40 1:0", "A:1323978162;C:446831209;G:581746343;T:2470114091;N:524086", 75, 0, null, null, 1323978162, 446831209, 581746343, 2470114091, 524086, "ERX4665134", "ERS5281175", "ERA3048543", "University of Zurich|European Nucleotide Archive", "University of Zurich|European Nucleotide Archive", 1, 0.36549, null, 0.19722, null, 0.95552, null, 0.4702, null, 76, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Switzerland", "2021-04-01", "Gastrula", "Embryo", "Embryo Imprecise", "All anatomical structures"], [10063, "ERR4795363", "ERX4665134", "ERS5281175", "ERP124847", "PRJEB41113", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E-MTAB-9727", "Transcriptome Analysis", "The mesothelium forms epithelial membranes that line the bodies cavities and surround the internal organs. Mesothelia widely contribute to organ homeostasis and regeneration  and their dysregulation can result in congenital anomalies of the viscera  ventral wall defects  and mesothelioma tumors. Nonetheless  the embryonic ontogeny and developmental regulation of mesothelium formation has remained uncharted. Here  we combine genetic lineage tracing  in toto live imaging  and single cell transcriptomics in zebrafish to track mesothelial progenitor origins from the lateral plate mesoderm LPM. Our single cell analysis uncovers a post gastrulation gene expression signature centered on hand2 that delineates distinct progenitor populations within the forming LPM. Combining gene expression analysis and imaging of transgenic reporter zebrafish embryos  we chart the origin of mesothelial progenitors to the lateral most  hand2 expressing LPM and confirm evolutionary conservation in mouse. Our time lapse imaging of transgenic hand2 reporter embryos captures zebrafish mesothelium formation  documenting the coordinated cell movements that form pericardium and visceral and parietal peritoneum. We establish that the primordial germ cells migrate associated with the forming mesothelium as ventral migration boundary. Functionally  hand2 mutants fail to close the ventral mesothelium due to perturbed migration of mesothelium progenitors. Analyzing mouse and human mesothelioma tumors hypothesized to emerge from transformed mesothelium  we find de novo expression of LPM associated transcription factors  and in particular of Hand2  indicating the re initiation of a developmental transcriptional program in mesothelioma. Taken together  our work outlines a genetic and developmental signature of mesothelial origins centered around Hand2  contributing to our understanding of mesothelial pathologies and mesothelioma.", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", null, "Protocols: Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Sample 3", "E MTAB 9727:Sample 3", null, "isolate:not applicable|organism:Danio rerio|sex:mixed|age:10|developmental stage:tailbud stage|organism part:lateral plate mesoderm plate|genotype:drl:mCherry|ENA FIRST PUBLIC:2021 04 01|ENA LAST UPDATE:2021 09 01", null, null, null, null, null, null, null, null, "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E MTAB 9727:Sample 3 s", "Sample 3 s", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Experimental Factor: replicate:lateral plate mesoderm plate 3", "RNA-Seq", "TRANSCRIPTOMIC SINGLE CELL", "RANDOM", "SINGLE", "ILLUMINA", "NextSeq 500", null, "ERP124847", "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", "SN3_AHY3WGBGX3_S7_R2_cat.fastq.gz", "fastq", 4828889391.0, 63965519.0, "E MTAB 9727:Sample 3 1", "0:0 1:75.49", "A:1434099697;C:964508763;G:893584575;T:1534727600;N:1968756", 0, 75, null, null, 1434099697, 964508763, 893584575, 1534727600, 1968756, "ERX4665134", "ERS5281175", "ERA3048543", "University of Zurich|European Nucleotide Archive", "University of Zurich|European Nucleotide Archive", 1, 0.85811, null, 0.26231, null, 0.82731, null, 0.48618, null, 76, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Switzerland", "2021-04-01", "Gastrula", "Embryo", "Embryo Imprecise", "All anatomical structures"], [10064, "ERR4795360", "ERX4665133", "ERS5281174", "ERP124847", "PRJEB41113", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E-MTAB-9727", "Transcriptome Analysis", "The mesothelium forms epithelial membranes that line the bodies cavities and surround the internal organs. Mesothelia widely contribute to organ homeostasis and regeneration  and their dysregulation can result in congenital anomalies of the viscera  ventral wall defects  and mesothelioma tumors. Nonetheless  the embryonic ontogeny and developmental regulation of mesothelium formation has remained uncharted. Here  we combine genetic lineage tracing  in toto live imaging  and single cell transcriptomics in zebrafish to track mesothelial progenitor origins from the lateral plate mesoderm LPM. Our single cell analysis uncovers a post gastrulation gene expression signature centered on hand2 that delineates distinct progenitor populations within the forming LPM. Combining gene expression analysis and imaging of transgenic reporter zebrafish embryos  we chart the origin of mesothelial progenitors to the lateral most  hand2 expressing LPM and confirm evolutionary conservation in mouse. Our time lapse imaging of transgenic hand2 reporter embryos captures zebrafish mesothelium formation  documenting the coordinated cell movements that form pericardium and visceral and parietal peritoneum. We establish that the primordial germ cells migrate associated with the forming mesothelium as ventral migration boundary. Functionally  hand2 mutants fail to close the ventral mesothelium due to perturbed migration of mesothelium progenitors. Analyzing mouse and human mesothelioma tumors hypothesized to emerge from transformed mesothelium  we find de novo expression of LPM associated transcription factors  and in particular of Hand2  indicating the re initiation of a developmental transcriptional program in mesothelioma. Taken together  our work outlines a genetic and developmental signature of mesothelial origins centered around Hand2  contributing to our understanding of mesothelial pathologies and mesothelioma.", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", null, "Protocols: Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Sample 2", "E MTAB 9727:Sample 2", null, "isolate:not applicable|organism:Danio rerio|sex:mixed|age:10|developmental stage:tailbud stage|organism part:lateral plate mesoderm plate|genotype:drl:mCherry|ENA FIRST PUBLIC:2021 04 01|ENA LAST UPDATE:2021 09 01", null, null, null, null, null, null, null, null, "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E MTAB 9727:Sample 2 s", "Sample 2 s", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Experimental Factor: replicate:lateral plate mesoderm plate 2", "RNA-Seq", "TRANSCRIPTOMIC SINGLE CELL", "RANDOM", "SINGLE", "ILLUMINA", "NextSeq 500", null, "ERP124847", "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", "SN2_AHY3WGBGX3_S6_R1_cat.fastq.gz", "fastq", 5598371393.0, 74235388.0, "E MTAB 9727:Sample 2", "0:75.41 1:0", "A:1526486973;C:558020193;G:717587491;T:2795646081;N:630655", 75, 0, null, null, 1526486973, 558020193, 717587491, 2795646081, 630655, "ERX4665133", "ERS5281174", "ERA3048543", "University of Zurich|European Nucleotide Archive", "University of Zurich|European Nucleotide Archive", 1, 0.28059, null, 0.19252, null, 0.96404, null, 0.4874, null, 75, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Switzerland", "2021-04-01", "Gastrula", "Embryo", "Embryo Imprecise", "All anatomical structures"], [10065, "ERR4795361", "ERX4665133", "ERS5281174", "ERP124847", "PRJEB41113", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E-MTAB-9727", "Transcriptome Analysis", "The mesothelium forms epithelial membranes that line the bodies cavities and surround the internal organs. Mesothelia widely contribute to organ homeostasis and regeneration  and their dysregulation can result in congenital anomalies of the viscera  ventral wall defects  and mesothelioma tumors. Nonetheless  the embryonic ontogeny and developmental regulation of mesothelium formation has remained uncharted. Here  we combine genetic lineage tracing  in toto live imaging  and single cell transcriptomics in zebrafish to track mesothelial progenitor origins from the lateral plate mesoderm LPM. Our single cell analysis uncovers a post gastrulation gene expression signature centered on hand2 that delineates distinct progenitor populations within the forming LPM. Combining gene expression analysis and imaging of transgenic reporter zebrafish embryos  we chart the origin of mesothelial progenitors to the lateral most  hand2 expressing LPM and confirm evolutionary conservation in mouse. Our time lapse imaging of transgenic hand2 reporter embryos captures zebrafish mesothelium formation  documenting the coordinated cell movements that form pericardium and visceral and parietal peritoneum. We establish that the primordial germ cells migrate associated with the forming mesothelium as ventral migration boundary. Functionally  hand2 mutants fail to close the ventral mesothelium due to perturbed migration of mesothelium progenitors. Analyzing mouse and human mesothelioma tumors hypothesized to emerge from transformed mesothelium  we find de novo expression of LPM associated transcription factors  and in particular of Hand2  indicating the re initiation of a developmental transcriptional program in mesothelioma. Taken together  our work outlines a genetic and developmental signature of mesothelial origins centered around Hand2  contributing to our understanding of mesothelial pathologies and mesothelioma.", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", null, "Protocols: Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Sample 2", "E MTAB 9727:Sample 2", null, "isolate:not applicable|organism:Danio rerio|sex:mixed|age:10|developmental stage:tailbud stage|organism part:lateral plate mesoderm plate|genotype:drl:mCherry|ENA FIRST PUBLIC:2021 04 01|ENA LAST UPDATE:2021 09 01", null, null, null, null, null, null, null, null, "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E MTAB 9727:Sample 2 s", "Sample 2 s", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Experimental Factor: replicate:lateral plate mesoderm plate 2", "RNA-Seq", "TRANSCRIPTOMIC SINGLE CELL", "RANDOM", "SINGLE", "ILLUMINA", "NextSeq 500", null, "ERP124847", "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", "SN2_AHY3WGBGX3_S6_R2_cat.fastq.gz", "fastq", 5600151371.0, 74235388.0, "E MTAB 9727:Sample 2 1", "0:0 1:75.44", "A:1850568176;C:1035622971;G:1071138281;T:1640475883;N:2346060", 0, 75, null, null, 1850568176, 1035622971, 1071138281, 1640475883, 2346060, "ERX4665133", "ERS5281174", "ERA3048543", "University of Zurich|European Nucleotide Archive", "University of Zurich|European Nucleotide Archive", 1, 0.7679, null, 0.30371, null, 0.82651, null, 0.43401, null, 75, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Switzerland", "2021-04-01", "Gastrula", "Embryo", "Embryo Imprecise", "All anatomical structures"], [10066, "ERR4795358", "ERX4665132", "ERS5281173", "ERP124847", "PRJEB41113", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E-MTAB-9727", "Transcriptome Analysis", "The mesothelium forms epithelial membranes that line the bodies cavities and surround the internal organs. Mesothelia widely contribute to organ homeostasis and regeneration  and their dysregulation can result in congenital anomalies of the viscera  ventral wall defects  and mesothelioma tumors. Nonetheless  the embryonic ontogeny and developmental regulation of mesothelium formation has remained uncharted. Here  we combine genetic lineage tracing  in toto live imaging  and single cell transcriptomics in zebrafish to track mesothelial progenitor origins from the lateral plate mesoderm LPM. Our single cell analysis uncovers a post gastrulation gene expression signature centered on hand2 that delineates distinct progenitor populations within the forming LPM. Combining gene expression analysis and imaging of transgenic reporter zebrafish embryos  we chart the origin of mesothelial progenitors to the lateral most  hand2 expressing LPM and confirm evolutionary conservation in mouse. Our time lapse imaging of transgenic hand2 reporter embryos captures zebrafish mesothelium formation  documenting the coordinated cell movements that form pericardium and visceral and parietal peritoneum. We establish that the primordial germ cells migrate associated with the forming mesothelium as ventral migration boundary. Functionally  hand2 mutants fail to close the ventral mesothelium due to perturbed migration of mesothelium progenitors. Analyzing mouse and human mesothelioma tumors hypothesized to emerge from transformed mesothelium  we find de novo expression of LPM associated transcription factors  and in particular of Hand2  indicating the re initiation of a developmental transcriptional program in mesothelioma. Taken together  our work outlines a genetic and developmental signature of mesothelial origins centered around Hand2  contributing to our understanding of mesothelial pathologies and mesothelioma.", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", null, "Protocols: Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Sample 1", "E MTAB 9727:Sample 1", null, "isolate:not applicable|organism:Danio rerio|sex:mixed|age:10|developmental stage:tailbud stage|organism part:lateral plate mesoderm plate|genotype:drl:mCherry|ENA FIRST PUBLIC:2021 04 01|ENA LAST UPDATE:2021 09 01", null, null, null, null, null, null, null, null, "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E MTAB 9727:Sample 1 s", "Sample 1 s", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Experimental Factor: replicate:lateral plate mesoderm plate 1", "RNA-Seq", "TRANSCRIPTOMIC SINGLE CELL", "RANDOM", "SINGLE", "ILLUMINA", "NextSeq 500", null, "ERP124847", "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", "SN1_AHY3WGBGX3_S5_R1_cat.fastq.gz", "fastq", 4996502316.0, 66258508.0, "E MTAB 9727:Sample 1", "0:75.41 1:0", "A:1330744174;C:451622042;G:591151500;T:2622422664;N:561936", 75, 0, null, null, 1330744174, 451622042, 591151500, 2622422664, 561936, "ERX4665132", "ERS5281173", "ERA3048543", "University of Zurich|European Nucleotide Archive", "University of Zurich|European Nucleotide Archive", 1, 0.29754, null, 0.17671, null, 0.9669, null, 0.45463, null, 75, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Switzerland", "2021-04-01", "Gastrula", "Embryo", "Embryo Imprecise", "All anatomical structures"], [10067, "ERR4795359", "ERX4665132", "ERS5281173", "ERP124847", "PRJEB41113", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E-MTAB-9727", "Transcriptome Analysis", "The mesothelium forms epithelial membranes that line the bodies cavities and surround the internal organs. Mesothelia widely contribute to organ homeostasis and regeneration  and their dysregulation can result in congenital anomalies of the viscera  ventral wall defects  and mesothelioma tumors. Nonetheless  the embryonic ontogeny and developmental regulation of mesothelium formation has remained uncharted. Here  we combine genetic lineage tracing  in toto live imaging  and single cell transcriptomics in zebrafish to track mesothelial progenitor origins from the lateral plate mesoderm LPM. Our single cell analysis uncovers a post gastrulation gene expression signature centered on hand2 that delineates distinct progenitor populations within the forming LPM. Combining gene expression analysis and imaging of transgenic reporter zebrafish embryos  we chart the origin of mesothelial progenitors to the lateral most  hand2 expressing LPM and confirm evolutionary conservation in mouse. Our time lapse imaging of transgenic hand2 reporter embryos captures zebrafish mesothelium formation  documenting the coordinated cell movements that form pericardium and visceral and parietal peritoneum. We establish that the primordial germ cells migrate associated with the forming mesothelium as ventral migration boundary. Functionally  hand2 mutants fail to close the ventral mesothelium due to perturbed migration of mesothelium progenitors. Analyzing mouse and human mesothelioma tumors hypothesized to emerge from transformed mesothelium  we find de novo expression of LPM associated transcription factors  and in particular of Hand2  indicating the re initiation of a developmental transcriptional program in mesothelioma. Taken together  our work outlines a genetic and developmental signature of mesothelial origins centered around Hand2  contributing to our understanding of mesothelial pathologies and mesothelioma.", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", null, "Protocols: Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Sample 1", "E MTAB 9727:Sample 1", null, "isolate:not applicable|organism:Danio rerio|sex:mixed|age:10|developmental stage:tailbud stage|organism part:lateral plate mesoderm plate|genotype:drl:mCherry|ENA FIRST PUBLIC:2021 04 01|ENA LAST UPDATE:2021 09 01", null, null, null, null, null, null, null, null, "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "E MTAB 9727:Sample 1 s", "Sample 1 s", "Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "Wildtype stain and drl:mCherry positive zebrafish embryos were grown until tailbud stage and chorions were removed by incubating in 1 mg/mL Pronase Roche  followed by washing in E3 medium. Around 300 embryos were used to generate the sample. The E3 medium was replaced by 1X PBS and the embryos were stored on ice until further processing. The embryos were dissociated using 2 mg/mL collagenase IV Worthington in DMEM high glucose 4.5g/l and NaHCO3  without xxx glutamine and sodium pyruvate  Sigma Aldrich and incubated for 5 min in a water bath at 37oC. The embryonic tissues were triturated into a single cell suspension by pipetting carefully up and down. When the embryos were not yet dissociated sufficiently  they were incubated for another 5 min. Cells were filtered through a 35 \u03bcm cell strainer Falcon        round bottom tubes with cell strainer cap and centrifuged at 6000 rpm for 30 sec. Cell pellets were resuspended in 1X HBSS Gibco containing 2% FBS and subjected to an additional round of centrifugation and resuspension. post washing  the cells were resuspended in 1X PBS.  drl:mCherry positive cells were sorted using a FACS Aria III cell sorter BD Bioscience. Cells were gated based on size and forward scattering  to exclude debris and doublets. The gating for the negative population was determined based on wildtype tailbud staged embryos. See also Figure S2 for gate setting. The SORTseq single cell RNA sequencing protocol was carried out as described previously Muraro et al.  2016. Live mCherry positive single cells were sorted in four 384 well plates BioRad containing 5 \u03bcl of CEL Seq2 primer solution in mineral oil 24 bp polyT stretch  a 4 bp random molecular barcode UMI  a cell specific barcode  the 5\u2032Illumina TruSeq small RNA kit adaptor and a T7 promoter  provided by Single Cell Discoveries. post sorting  the plates were immediately placed on ice and stored at \u221280\u00b0C.  In brief  ERCC Spike in RNA 0.02 \u03bcL of 1:50000 dilution was added to each well before cell lysis with heat shocking. Reverse transcription and second strand synthesis reagents were dispensed using the Nanodrop II GC biotech. post generation of cDNA from the original mRNA  all cells from one plate were pooled and the pooled sample was amplified linearly with in vitro transcription. The amplified RNA was then reverse transcribed to cDNA using a random hexamer primers Hashimshony et al.  2016. To generate sequencing libraries  RPI series index primers were used for library PCR.", "Experimental Factor: replicate:lateral plate mesoderm plate 1", "RNA-Seq", "TRANSCRIPTOMIC SINGLE CELL", "RANDOM", "SINGLE", "ILLUMINA", "NextSeq 500", null, "ERP124847", "NextSeq 500 sequencing; Hand2 delineates mesothelium progenitors and is reactivated in mesothelioma", "ENA FIRST PUBLIC:2022 01 14|ENA LAST UPDATE:2022 01 14", "SN1_AHY3WGBGX3_S5_R2_cat.fastq.gz", "fastq", 4999908389.0, 66258508.0, "E MTAB 9727:Sample 1 1", "0:0 1:75.46", "A:1591945117;C:933968124;G:986108614;T:1485824022;N:2062512", 0, 75, null, null, 1591945117, 933968124, 986108614, 1485824022, 2062512, "ERX4665132", "ERS5281173", "ERA3048543", "University of Zurich|European Nucleotide Archive", "University of Zurich|European Nucleotide Archive", 1, 0.81333, null, 0.2466, null, 0.83027, null, 0.51572, null, 75, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Switzerland", "2021-04-01", "Gastrula", "Embryo", "Embryo Imprecise", "All anatomical structures"], [30508, "SRR27764648", "SRX23429609", "SRS20284060", "SRP486518", "PRJNA1070582", "Gene expression profile of neighboring cells of cells with unfit Wnt morphogen gradient during cell competition", "GSE254439", "Transcriptome Analysis", "Morphogen signalling forms an activity gradient and instructs cell identities in a signalling strength dependent manner to pattern developing tissues. However  developing tissues also undergo dynamic morphogenesis  which may produce cells with unfit morphogen signalling and consequent noisy morphogen gradient. Here we show that a cell competition related system corrects such noisy morphogen gradients. Zebrafish imaging analyses of the Wnt/\u00df catenin signalling gradient  which acts as a morphogen to establish embryonic anterior posterior patterning  revealed that unfit cells with abnormal Wnt/\u00df catenin activity spontaneously appear and produce noise in the gradient. Communication between unfit and neighbouring fit cells via cadherin proteins stimulates apoptosis of the unfit cells by activating Smad signalling and reactive oxygen species production. This unfit cell elimination is required for proper Wnt/\u00df catenin gradient formation and consequent anterior posterior patterning. Because this gradient controls patterning not only in the embryo but also in adult tissues  this system may support tissue robustness and disease prevention. Overall design: \u00df catCA constitutive active form of \u00df catenin expressing GFP+ cells  or \u00dfcatCA unexpressing GFP  cells from \u00df catCA mosaically introduced zebrafish early embryos were sorted by FACS cell sorter. Total RNAs were extracted and analyzed by RNA seq.", null, "pubmed:39546611", null, "Mosaic GFP control GFP min lot2", "GSM8042248", null, "source name:early embryonic cell|strain:AB|tissue:early embryonic cell|developmental stage:9 hpf loc name:missing|collection date:missing", "Mosaic GFP control GFP min lot2", "Sequenced reads were mapped to the GRCz10 reference genome using Bowtie2 ver. 2.3.1. UMI counts were extracted per HTSeq ver. 0.6.1 from the CEL Seq2 pipeline. Assembly: GRCz10 Supplementary files format and content: Tab delimited text file for expression data normalized by regularized logarithm.", "early embryonic cell", null, "Cells were dissolved in TRIzol reagent Invitrogen and extracted. Samples were prepared according to the CEL Seq2 protocol described in Hashimshony et al. Geneome Biology 2016. Libraries preparation was performed according to the published CEL Seq2 protocol.", null, "strain:AB|tissue:early embryonic cell|developmental stage:9 hpf", "GSM8042248", "GSM8042248: Mosaic GFP control GFP min lot2; Danio rerio; RNA Seq", "GSM8042248 r1", "GSM8042248", "1", "Cells were dissolved in TRIzol reagent Invitrogen and extracted. Samples were prepared according to the CEL Seq2 protocol described in Hashimshony et al. Geneome Biology 2016. Libraries preparation was performed according to the published CEL Seq2 protocol.", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 1500", null, "SRP486518", null, null, "Mosaic_GFP_control_GFP_min_lot2.fastq.gz", "fastq", 7474464.0, 207624.0, "GSM8042248 r1", "0:36", "A:1796571;C:1274347;G:1548876;T:2854637;N:33", 36, null, null, null, 1796571, 1274347, 1548876, 2854637, 33, "SRX23429609", "SRS20284060", "SRA1793856", "Department of Homeostatic Regulation, Research Institute for Microbial Diseases, Osaka University", "Department of Homeostatic Regulation, Research Institute for Microbial Diseases, Osaka University", 1, 0.82922, null, 0.20207, null, 0.86143, null, 0.54661, null, 36, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Japan", "2024-01-29", "Gastrula", "Embryo", "Embryo Imprecise", "All anatomical structures"], [30509, "SRR27764649", "SRX23429608", "SRS20284059", "SRP486518", "PRJNA1070582", "Gene expression profile of neighboring cells of cells with unfit Wnt morphogen gradient during cell competition", "GSE254439", "Transcriptome Analysis", "Morphogen signalling forms an activity gradient and instructs cell identities in a signalling strength dependent manner to pattern developing tissues. However  developing tissues also undergo dynamic morphogenesis  which may produce cells with unfit morphogen signalling and consequent noisy morphogen gradient. Here we show that a cell competition related system corrects such noisy morphogen gradients. Zebrafish imaging analyses of the Wnt/\u00df catenin signalling gradient  which acts as a morphogen to establish embryonic anterior posterior patterning  revealed that unfit cells with abnormal Wnt/\u00df catenin activity spontaneously appear and produce noise in the gradient. Communication between unfit and neighbouring fit cells via cadherin proteins stimulates apoptosis of the unfit cells by activating Smad signalling and reactive oxygen species production. This unfit cell elimination is required for proper Wnt/\u00df catenin gradient formation and consequent anterior posterior patterning. Because this gradient controls patterning not only in the embryo but also in adult tissues  this system may support tissue robustness and disease prevention. Overall design: \u00df catCA constitutive active form of \u00df catenin expressing GFP+ cells  or \u00dfcatCA unexpressing GFP  cells from \u00df catCA mosaically introduced zebrafish early embryos were sorted by FACS cell sorter. Total RNAs were extracted and analyzed by RNA seq.", null, "pubmed:39546611", null, "Mosaic GFP control GFP min lot1", "GSM8042247", null, "source name:early embryonic cell|strain:AB|tissue:early embryonic cell|developmental stage:9 hpf loc name:missing|collection date:missing", "Mosaic GFP control GFP min lot1", "Sequenced reads were mapped to the GRCz10 reference genome using Bowtie2 ver. 2.3.1. UMI counts were extracted per HTSeq ver. 0.6.1 from the CEL Seq2 pipeline. Assembly: GRCz10 Supplementary files format and content: Tab delimited text file for expression data normalized by regularized logarithm.", "early embryonic cell", null, "Cells were dissolved in TRIzol reagent Invitrogen and extracted. Samples were prepared according to the CEL Seq2 protocol described in Hashimshony et al. Geneome Biology 2016. Libraries preparation was performed according to the published CEL Seq2 protocol.", null, "strain:AB|tissue:early embryonic cell|developmental stage:9 hpf", "GSM8042247", "GSM8042247: Mosaic GFP control GFP min lot1; Danio rerio; RNA Seq", "GSM8042247 r1", "GSM8042247", "1", "Cells were dissolved in TRIzol reagent Invitrogen and extracted. Samples were prepared according to the CEL Seq2 protocol described in Hashimshony et al. Geneome Biology 2016. Libraries preparation was performed according to the published CEL Seq2 protocol.", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 1500", null, "SRP486518", null, null, "Mosaic_GFP_control_GFP_min_lot1.fastq.gz", "fastq", 8256240.0, 229340.0, "GSM8042247 r1", "0:36", "A:1947003;C:1345938;G:1692065;T:3271133;N:101", 36, null, null, null, 1947003, 1345938, 1692065, 3271133, 101, "SRX23429608", "SRS20284059", "SRA1793856", "Department of Homeostatic Regulation, Research Institute for Microbial Diseases, Osaka University", "Department of Homeostatic Regulation, Research Institute for Microbial Diseases, Osaka University", 1, 0.83305, null, 0.17754, null, 0.8619, null, 0.61474, null, 36, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Japan", "2024-01-29", "Gastrula", "Embryo", "Embryo Imprecise", "All anatomical structures"], [30510, "SRR27764650", "SRX23429607", "SRS20284058", "SRP486518", "PRJNA1070582", "Gene expression profile of neighboring cells of cells with unfit Wnt morphogen gradient during cell competition", "GSE254439", "Transcriptome Analysis", "Morphogen signalling forms an activity gradient and instructs cell identities in a signalling strength dependent manner to pattern developing tissues. However  developing tissues also undergo dynamic morphogenesis  which may produce cells with unfit morphogen signalling and consequent noisy morphogen gradient. Here we show that a cell competition related system corrects such noisy morphogen gradients. Zebrafish imaging analyses of the Wnt/\u00df catenin signalling gradient  which acts as a morphogen to establish embryonic anterior posterior patterning  revealed that unfit cells with abnormal Wnt/\u00df catenin activity spontaneously appear and produce noise in the gradient. Communication between unfit and neighbouring fit cells via cadherin proteins stimulates apoptosis of the unfit cells by activating Smad signalling and reactive oxygen species production. This unfit cell elimination is required for proper Wnt/\u00df catenin gradient formation and consequent anterior posterior patterning. Because this gradient controls patterning not only in the embryo but also in adult tissues  this system may support tissue robustness and disease prevention. Overall design: \u00df catCA constitutive active form of \u00df catenin expressing GFP+ cells  or \u00dfcatCA unexpressing GFP  cells from \u00df catCA mosaically introduced zebrafish early embryos were sorted by FACS cell sorter. Total RNAs were extracted and analyzed by RNA seq.", null, "pubmed:39546611", null, "Mosaic beta catCA GFP min lot2", "GSM8042246", null, "source name:early embryonic cell|strain:AB|tissue:early embryonic cell|developmental stage:9 hpf loc name:missing|collection date:missing", "Mosaic beta catCA GFP min lot2", "Sequenced reads were mapped to the GRCz10 reference genome using Bowtie2 ver. 2.3.1. UMI counts were extracted per HTSeq ver. 0.6.1 from the CEL Seq2 pipeline. Assembly: GRCz10 Supplementary files format and content: Tab delimited text file for expression data normalized by regularized logarithm.", "early embryonic cell", null, "Cells were dissolved in TRIzol reagent Invitrogen and extracted. Samples were prepared according to the CEL Seq2 protocol described in Hashimshony et al. Geneome Biology 2016. Libraries preparation was performed according to the published CEL Seq2 protocol.", null, "strain:AB|tissue:early embryonic cell|developmental stage:9 hpf", "GSM8042246", "GSM8042246: Mosaic beta catCA GFP min lot2; Danio rerio; RNA Seq", "GSM8042246 r1", "GSM8042246", "1", "Cells were dissolved in TRIzol reagent Invitrogen and extracted. Samples were prepared according to the CEL Seq2 protocol described in Hashimshony et al. Geneome Biology 2016. Libraries preparation was performed according to the published CEL Seq2 protocol.", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 1500", null, "SRP486518", null, null, "Mosaic_beta-catCA_GFP_min_lot2.fastq.gz", "fastq", 856836.0, 23801.0, "GSM8042246 r1", "0:36", "A:217589;C:147419;G:175612;T:316201;N:15", 36, null, null, null, 217589, 147419, 175612, 316201, 15, "SRX23429607", "SRS20284058", "SRA1793856", "Department of Homeostatic Regulation, Research Institute for Microbial Diseases, Osaka University", "Department of Homeostatic Regulation, Research Institute for Microbial Diseases, Osaka University", 1, 0.82481, null, 0.28434, null, 0.95371, null, 0.61551, null, 36, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Japan", "2024-01-29", "Gastrula", "Embryo", "Embryo Imprecise", "All anatomical structures"], [30511, "SRR27764651", "SRX23429606", "SRS20284057", "SRP486518", "PRJNA1070582", "Gene expression profile of neighboring cells of cells with unfit Wnt morphogen gradient during cell competition", "GSE254439", "Transcriptome Analysis", "Morphogen signalling forms an activity gradient and instructs cell identities in a signalling strength dependent manner to pattern developing tissues. However  developing tissues also undergo dynamic morphogenesis  which may produce cells with unfit morphogen signalling and consequent noisy morphogen gradient. Here we show that a cell competition related system corrects such noisy morphogen gradients. Zebrafish imaging analyses of the Wnt/\u00df catenin signalling gradient  which acts as a morphogen to establish embryonic anterior posterior patterning  revealed that unfit cells with abnormal Wnt/\u00df catenin activity spontaneously appear and produce noise in the gradient. Communication between unfit and neighbouring fit cells via cadherin proteins stimulates apoptosis of the unfit cells by activating Smad signalling and reactive oxygen species production. This unfit cell elimination is required for proper Wnt/\u00df catenin gradient formation and consequent anterior posterior patterning. Because this gradient controls patterning not only in the embryo but also in adult tissues  this system may support tissue robustness and disease prevention. Overall design: \u00df catCA constitutive active form of \u00df catenin expressing GFP+ cells  or \u00dfcatCA unexpressing GFP  cells from \u00df catCA mosaically introduced zebrafish early embryos were sorted by FACS cell sorter. Total RNAs were extracted and analyzed by RNA seq.", null, "pubmed:39546611", null, "Mosaic beta catCA GFP min lot1", "GSM8042245", null, "source name:early embryonic cell|strain:AB|tissue:early embryonic cell|developmental stage:9 hpf loc name:missing|collection date:missing", "Mosaic beta catCA GFP min lot1", "Sequenced reads were mapped to the GRCz10 reference genome using Bowtie2 ver. 2.3.1. UMI counts were extracted per HTSeq ver. 0.6.1 from the CEL Seq2 pipeline. Assembly: GRCz10 Supplementary files format and content: Tab delimited text file for expression data normalized by regularized logarithm.", "early embryonic cell", null, "Cells were dissolved in TRIzol reagent Invitrogen and extracted. Samples were prepared according to the CEL Seq2 protocol described in Hashimshony et al. Geneome Biology 2016. Libraries preparation was performed according to the published CEL Seq2 protocol.", null, "strain:AB|tissue:early embryonic cell|developmental stage:9 hpf", "GSM8042245", "GSM8042245: Mosaic beta catCA GFP min lot1; Danio rerio; RNA Seq", "GSM8042245 r1", "GSM8042245", "1", "Cells were dissolved in TRIzol reagent Invitrogen and extracted. Samples were prepared according to the CEL Seq2 protocol described in Hashimshony et al. Geneome Biology 2016. Libraries preparation was performed according to the published CEL Seq2 protocol.", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 1500", null, "SRP486518", null, null, "Mosaic_beta-catCA_GFP_min_lot1.fastq.gz", "fastq", 7181388.0, 199483.0, "GSM8042245 r1", "0:36", "A:1744919;C:1207643;G:1456829;T:2771919;N:78", 36, null, null, null, 1744919, 1207643, 1456829, 2771919, 78, "SRX23429606", "SRS20284057", "SRA1793856", "Department of Homeostatic Regulation, Research Institute for Microbial Diseases, Osaka University", "Department of Homeostatic Regulation, Research Institute for Microbial Diseases, Osaka University", 1, 0.83629, null, 0.16823, null, 0.84774, null, 0.64548, null, 36, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Japan", "2024-01-29", "Gastrula", "Embryo", "Embryo Imprecise", "All anatomical structures"], [33075, "SRR29654109", "SRX25158199", "SRS21848821", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 3", "GSM8369980", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100", "GSM8369980", "GSM8369980: Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 3; Danio rerio; RNA Seq", "GSM8369980 r1", "GSM8369980", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L001_8hpi_AMD_-016_cbc.fastq.gz", "fastq", 347162880.0, 5786048.0, "GSM8369980 r1", "0:60", "A:122562825;C:62219285;G:75539895;T:86775172;N:65703", 60, null, null, null, 122562825, 62219285, 75539895, 86775172, 65703, "SRX25158199", "SRS21848821", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33076, "SRR29654110", "SRX25158199", "SRS21848821", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 3", "GSM8369980", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100", "GSM8369980", "GSM8369980: Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 3; Danio rerio; RNA Seq", "GSM8369980 r1", "GSM8369980", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L002_8hpi_AMD_-016_cbc.fastq.gz", "fastq", 340954260.0, 5682571.0, "GSM8369980 r2", "0:60", "A:120149623;C:60854408;G:75192893;T:84703966;N:53370", 60, null, null, null, 120149623, 60854408, 75192893, 84703966, 53370, "SRX25158199", "SRS21848821", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33077, "SRR29654111", "SRX25158199", "SRS21848821", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 3", "GSM8369980", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100", "GSM8369980", "GSM8369980: Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 3; Danio rerio; RNA Seq", "GSM8369980 r1", "GSM8369980", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L003_8hpi_AMD_-016_cbc.fastq.gz", "fastq", 351749160.0, 5862486.0, "GSM8369980 r3", "0:60", "A:124352905;C:63078724;G:76291828;T:87988386;N:37317", 60, null, null, null, 124352905, 63078724, 76291828, 87988386, 37317, "SRX25158199", "SRS21848821", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33078, "SRR29654112", "SRX25158199", "SRS21848821", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 3", "GSM8369980", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100", "GSM8369980", "GSM8369980: Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 3; Danio rerio; RNA Seq", "GSM8369980 r1", "GSM8369980", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L004_8hpi_AMD_-016_cbc.fastq.gz", "fastq", 345225720.0, 5753762.0, "GSM8369980 r4", "0:60", "A:121803902;C:61651407;G:75900430;T:85843789;N:26192", 60, null, null, null, 121803902, 61651407, 75900430, 85843789, 26192, "SRX25158199", "SRS21848821", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33079, "SRR29654113", "SRX25158198", "SRS21848820", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 2", "GSM8369979", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100", "GSM8369979", "GSM8369979: Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 2; Danio rerio; RNA Seq", "GSM8369979 r1", "GSM8369979", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L001_8hpi_AMD_-015_cbc.fastq.gz", "fastq", 159497640.0, 2658294.0, "GSM8369979 r1", "0:60", "A:55569742;C:28678683;G:33264744;T:41953354;N:31117", 60, null, null, null, 55569742, 28678683, 33264744, 41953354, 31117, "SRX25158198", "SRS21848820", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33080, "SRR29654114", "SRX25158198", "SRS21848820", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 2", "GSM8369979", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100", "GSM8369979", "GSM8369979: Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 2; Danio rerio; RNA Seq", "GSM8369979 r1", "GSM8369979", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L002_8hpi_AMD_-015_cbc.fastq.gz", "fastq", 156549240.0, 2609154.0, "GSM8369979 r2", "0:60", "A:54428858;C:28027289;G:33149650;T:40918247;N:25196", 60, null, null, null, 54428858, 28027289, 33149650, 40918247, 25196, "SRX25158198", "SRS21848820", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33081, "SRR29654115", "SRX25158198", "SRS21848820", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 2", "GSM8369979", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100", "GSM8369979", "GSM8369979: Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 2; Danio rerio; RNA Seq", "GSM8369979 r1", "GSM8369979", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L003_8hpi_AMD_-015_cbc.fastq.gz", "fastq", 161735280.0, 2695588.0, "GSM8369979 r3", "0:60", "A:56425187;C:29084972;G:33637112;T:42570603;N:17406", 60, null, null, null, 56425187, 29084972, 33637112, 42570603, 17406, "SRX25158198", "SRS21848820", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33082, "SRR29654116", "SRX25158198", "SRS21848820", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 2", "GSM8369979", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100", "GSM8369979", "GSM8369979: Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 2; Danio rerio; RNA Seq", "GSM8369979 r1", "GSM8369979", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L004_8hpi_AMD_-015_cbc.fastq.gz", "fastq", 158621520.0, 2643692.0, "GSM8369979 r4", "0:60", "A:55213721;C:28430013;G:33462307;T:41503213;N:12266", 60, null, null, null, 55213721, 28430013, 33462307, 41503213, 12266, "SRX25158198", "SRS21848820", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33083, "SRR29654117", "SRX25158197", "SRS21848819", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 1", "GSM8369978", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100", "GSM8369978", "GSM8369978: Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 1; Danio rerio; RNA Seq", "GSM8369978 r1", "GSM8369978", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L001_8hpi_AMD_-011_cbc.fastq.gz", "fastq", 44363040.0, 739384.0, "GSM8369978 r1", "0:60", "A:17756825;C:7540147;G:9311367;T:9746444;N:8257", 60, null, null, null, 17756825, 7540147, 9311367, 9746444, 8257, "SRX25158197", "SRS21848819", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33084, "SRR29654118", "SRX25158197", "SRS21848819", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 1", "GSM8369978", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100", "GSM8369978", "GSM8369978: Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 1; Danio rerio; RNA Seq", "GSM8369978 r1", "GSM8369978", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L002_8hpi_AMD_-011_cbc.fastq.gz", "fastq", 43450800.0, 724180.0, "GSM8369978 r2", "0:60", "A:17376442;C:7355139;G:9244019;T:9468455;N:6745", 60, null, null, null, 17376442, 7355139, 9244019, 9468455, 6745, "SRX25158197", "SRS21848819", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33085, "SRR29654119", "SRX25158197", "SRS21848819", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 1", "GSM8369978", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100", "GSM8369978", "GSM8369978: Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 1; Danio rerio; RNA Seq", "GSM8369978 r1", "GSM8369978", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L003_8hpi_AMD_-011_cbc.fastq.gz", "fastq", 44795880.0, 746598.0, "GSM8369978 r3", "0:60", "A:17988707;C:7600964;G:9362568;T:9839285;N:4356", 60, null, null, null, 17988707, 7600964, 9362568, 9839285, 4356, "SRX25158197", "SRS21848819", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33086, "SRR29654120", "SRX25158197", "SRS21848819", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 1", "GSM8369978", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury AMD3100", "GSM8369978", "GSM8369978: Neutrophils mpx+  8 xxx post injury  AMD3100  biol rep 1; Danio rerio; RNA Seq", "GSM8369978 r1", "GSM8369978", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L004_8hpi_AMD_-011_cbc.fastq.gz", "fastq", 44060940.0, 734349.0, "GSM8369978 r4", "0:60", "A:17608648;C:7492643;G:9327676;T:9628642;N:3331", 60, null, null, null, 17608648, 7492643, 9327676, 9628642, 3331, "SRX25158197", "SRS21848819", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33087, "SRR29654121", "SRX25158196", "SRS21848818", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  AMD3100  biol rep 3", "GSM8369977", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  AMD3100  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100", "GSM8369977", "GSM8369977: Neutrophils mpx+  uninjured  AMD3100  biol rep 3; Danio rerio; RNA Seq", "GSM8369977 r1", "GSM8369977", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L001_Uninjured_AMD_-014_cbc.fastq.gz", "fastq", 115048140.0, 1917469.0, "GSM8369977 r1", "0:60", "A:41373215;C:21759509;G:21832537;T:30061744;N:21135", 60, null, null, null, 41373215, 21759509, 21832537, 30061744, 21135, "SRX25158196", "SRS21848818", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33088, "SRR29654122", "SRX25158196", "SRS21848818", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  AMD3100  biol rep 3", "GSM8369977", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  AMD3100  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100", "GSM8369977", "GSM8369977: Neutrophils mpx+  uninjured  AMD3100  biol rep 3; Danio rerio; RNA Seq", "GSM8369977 r1", "GSM8369977", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L002_Uninjured_AMD_-014_cbc.fastq.gz", "fastq", 112845060.0, 1880751.0, "GSM8369977 r2", "0:60", "A:40482360;C:21287713;G:21750928;T:29306465;N:17594", 60, null, null, null, 40482360, 21287713, 21750928, 29306465, 17594, "SRX25158196", "SRS21848818", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33089, "SRR29654123", "SRX25158196", "SRS21848818", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  AMD3100  biol rep 3", "GSM8369977", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  AMD3100  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100", "GSM8369977", "GSM8369977: Neutrophils mpx+  uninjured  AMD3100  biol rep 3; Danio rerio; RNA Seq", "GSM8369977 r1", "GSM8369977", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L003_Uninjured_AMD_-014_cbc.fastq.gz", "fastq", 116696580.0, 1944943.0, "GSM8369977 r3", "0:60", "A:42022776;C:22106040;G:22075778;T:30479513;N:12473", 60, null, null, null, 42022776, 22106040, 22075778, 30479513, 12473, "SRX25158196", "SRS21848818", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33090, "SRR29654124", "SRX25158196", "SRS21848818", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  AMD3100  biol rep 3", "GSM8369977", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  AMD3100  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100", "GSM8369977", "GSM8369977: Neutrophils mpx+  uninjured  AMD3100  biol rep 3; Danio rerio; RNA Seq", "GSM8369977 r1", "GSM8369977", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L004_Uninjured_AMD_-014_cbc.fastq.gz", "fastq", 114334980.0, 1905583.0, "GSM8369977 r4", "0:60", "A:41132105;C:21578779;G:21934073;T:29681512;N:8511", 60, null, null, null, 41132105, 21578779, 21934073, 29681512, 8511, "SRX25158196", "SRS21848818", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33091, "SRR29654125", "SRX25158195", "SRS21848817", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  AMD3100  biol rep 2", "GSM8369976", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  AMD3100  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100", "GSM8369976", "GSM8369976: Neutrophils mpx+  uninjured  AMD3100  biol rep 2; Danio rerio; RNA Seq", "GSM8369976 r1", "GSM8369976", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L001_Uninjured_AMD_-013_cbc.fastq.gz", "fastq", 354085380.0, 5901423.0, "GSM8369976 r1", "0:60", "A:124800871;C:65627966;G:72434440;T:91152029;N:70074", 60, null, null, null, 124800871, 65627966, 72434440, 91152029, 70074, "SRX25158195", "SRS21848817", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33092, "SRR29654126", "SRX25158195", "SRS21848817", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  AMD3100  biol rep 2", "GSM8369976", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  AMD3100  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100", "GSM8369976", "GSM8369976: Neutrophils mpx+  uninjured  AMD3100  biol rep 2; Danio rerio; RNA Seq", "GSM8369976 r1", "GSM8369976", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L002_Uninjured_AMD_-013_cbc.fastq.gz", "fastq", 347250360.0, 5787506.0, "GSM8369976 r2", "0:60", "A:122147080;C:64042522;G:72135325;T:88869927;N:55506", 60, null, null, null, 122147080, 64042522, 72135325, 88869927, 55506, "SRX25158195", "SRS21848817", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33093, "SRR29654127", "SRX25158195", "SRS21848817", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  AMD3100  biol rep 2", "GSM8369976", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  AMD3100  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100", "GSM8369976", "GSM8369976: Neutrophils mpx+  uninjured  AMD3100  biol rep 2; Danio rerio; RNA Seq", "GSM8369976 r1", "GSM8369976", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L003_Uninjured_AMD_-013_cbc.fastq.gz", "fastq", 358862580.0, 5981043.0, "GSM8369976 r3", "0:60", "A:126636264;C:66527902;G:73214426;T:92445348;N:38640", 60, null, null, null, 126636264, 66527902, 73214426, 92445348, 38640, "SRX25158195", "SRS21848817", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33094, "SRR29654128", "SRX25158195", "SRS21848817", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  AMD3100  biol rep 2", "GSM8369976", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  AMD3100  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100", "GSM8369976", "GSM8369976: Neutrophils mpx+  uninjured  AMD3100  biol rep 2; Danio rerio; RNA Seq", "GSM8369976 r1", "GSM8369976", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L004_Uninjured_AMD_-013_cbc.fastq.gz", "fastq", 351843120.0, 5864052.0, "GSM8369976 r4", "0:60", "A:123921807;C:64956560;G:72828989;T:90107944;N:27820", 60, null, null, null, 123921807, 64956560, 72828989, 90107944, 27820, "SRX25158195", "SRS21848817", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33095, "SRR29654129", "SRX25158194", "SRS21848816", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  AMD3100  biol rep 1", "GSM8369975", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  AMD3100  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100", "GSM8369975", "GSM8369975: Neutrophils mpx+  uninjured  AMD3100  biol rep 1; Danio rerio; RNA Seq", "GSM8369975 r1", "GSM8369975", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L001_Uninjured_AMD_-010_cbc.fastq.gz", "fastq", 100732380.0, 1678873.0, "GSM8369975 r1", "0:60", "A:37215898;C:18610797;G:18111317;T:26774380;N:19988", 60, null, null, null, 37215898, 18610797, 18111317, 26774380, 19988, "SRX25158194", "SRS21848816", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33096, "SRR29654130", "SRX25158194", "SRS21848816", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  AMD3100  biol rep 1", "GSM8369975", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  AMD3100  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100", "GSM8369975", "GSM8369975: Neutrophils mpx+  uninjured  AMD3100  biol rep 1; Danio rerio; RNA Seq", "GSM8369975 r1", "GSM8369975", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L002_Uninjured_AMD_-010_cbc.fastq.gz", "fastq", 98830860.0, 1647181.0, "GSM8369975 r2", "0:60", "A:36388012;C:18195984;G:18122717;T:26108692;N:15455", 60, null, null, null, 36388012, 18195984, 18122717, 26108692, 15455, "SRX25158194", "SRS21848816", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33097, "SRR29654131", "SRX25158194", "SRS21848816", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  AMD3100  biol rep 1", "GSM8369975", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  AMD3100  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100", "GSM8369975", "GSM8369975: Neutrophils mpx+  uninjured  AMD3100  biol rep 1; Danio rerio; RNA Seq", "GSM8369975 r1", "GSM8369975", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L003_Uninjured_AMD_-010_cbc.fastq.gz", "fastq", 102277560.0, 1704626.0, "GSM8369975 r3", "0:60", "A:37811843;C:18928051;G:18347174;T:27179477;N:11015", 60, null, null, null, 37811843, 18928051, 18347174, 27179477, 11015, "SRX25158194", "SRS21848816", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33098, "SRR29654132", "SRX25158194", "SRS21848816", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  AMD3100  biol rep 1", "GSM8369975", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  AMD3100  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured AMD3100", "GSM8369975", "GSM8369975: Neutrophils mpx+  uninjured  AMD3100  biol rep 1; Danio rerio; RNA Seq", "GSM8369975 r1", "GSM8369975", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L004_Uninjured_AMD_-010_cbc.fastq.gz", "fastq", 100371840.0, 1672864.0, "GSM8369975 r4", "0:60", "A:37032383;C:18500721;G:18308236;T:26522753;N:7747", 60, null, null, null, 37032383, 18500721, 18308236, 26522753, 7747, "SRX25158194", "SRS21848816", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33099, "SRR29654133", "SRX25158193", "SRS21848815", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 3", "GSM8369974", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO", "GSM8369974", "GSM8369974: Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 3; Danio rerio; RNA Seq", "GSM8369974 r1", "GSM8369974", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L001_8hpi_DMSO_-008_cbc.fastq.gz", "fastq", 161717520.0, 2695292.0, "GSM8369974 r1", "0:60", "A:58227035;C:29120167;G:33153372;T:41186362;N:30584", 60, null, null, null, 58227035, 29120167, 33153372, 41186362, 30584, "SRX25158193", "SRS21848815", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33100, "SRR29654134", "SRX25158193", "SRS21848815", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 3", "GSM8369974", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO", "GSM8369974", "GSM8369974: Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 3; Danio rerio; RNA Seq", "GSM8369974 r1", "GSM8369974", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L002_8hpi_DMSO_-008_cbc.fastq.gz", "fastq", 158810460.0, 2646841.0, "GSM8369974 r2", "0:60", "A:57059029;C:28475827;G:33090143;T:40160441;N:25020", 60, null, null, null, 57059029, 28475827, 33090143, 40160441, 25020, "SRX25158193", "SRS21848815", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33101, "SRR29654135", "SRX25158193", "SRS21848815", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 3", "GSM8369974", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO", "GSM8369974", "GSM8369974: Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 3; Danio rerio; RNA Seq", "GSM8369974 r1", "GSM8369974", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L003_8hpi_DMSO_-008_cbc.fastq.gz", "fastq", 164136600.0, 2735610.0, "GSM8369974 r3", "0:60", "A:59164760;C:29564845;G:33567677;T:41822336;N:16982", 60, null, null, null, 59164760, 29564845, 33567677, 41822336, 16982, "SRX25158193", "SRS21848815", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33102, "SRR29654136", "SRX25158193", "SRS21848815", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 3", "GSM8369974", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO", "GSM8369974", "GSM8369974: Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 3; Danio rerio; RNA Seq", "GSM8369974 r1", "GSM8369974", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L004_8hpi_DMSO_-008_cbc.fastq.gz", "fastq", 160939740.0, 2682329.0, "GSM8369974 r4", "0:60", "A:57905646;C:28872433;G:33402085;T:40746636;N:12940", 60, null, null, null, 57905646, 28872433, 33402085, 40746636, 12940, "SRX25158193", "SRS21848815", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33103, "SRR29654137", "SRX25158192", "SRS21848814", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 2", "GSM8369973", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO", "GSM8369973", "GSM8369973: Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 2; Danio rerio; RNA Seq", "GSM8369973 r1", "GSM8369973", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L001_8hpi_DMSO_-005_cbc.fastq.gz", "fastq", 294896760.0, 4914946.0, "GSM8369973 r1", "0:60", "A:113302890;C:53832811;G:54274745;T:73429183;N:57131", 60, null, null, null, 113302890, 53832811, 54274745, 73429183, 57131, "SRX25158192", "SRS21848814", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33104, "SRR29654138", "SRX25158192", "SRS21848814", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 2", "GSM8369973", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO", "GSM8369973", "GSM8369973: Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 2; Danio rerio; RNA Seq", "GSM8369973 r1", "GSM8369973", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L002_8hpi_DMSO_-005_cbc.fastq.gz", "fastq", 289658820.0, 4827647.0, "GSM8369973 r2", "0:60", "A:111138289;C:52646716;G:54190664;T:71637241;N:45910", 60, null, null, null, 111138289, 52646716, 54190664, 71637241, 45910, "SRX25158192", "SRS21848814", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33105, "SRR29654139", "SRX25158192", "SRS21848814", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 2", "GSM8369973", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO", "GSM8369973", "GSM8369973: Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 2; Danio rerio; RNA Seq", "GSM8369973 r1", "GSM8369973", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L003_8hpi_DMSO_-005_cbc.fastq.gz", "fastq", 299290800.0, 4988180.0, "GSM8369973 r3", "0:60", "A:115247052;C:54662821;G:54854607;T:74494831;N:31489", 60, null, null, null, 115247052, 54662821, 54854607, 74494831, 31489, "SRX25158192", "SRS21848814", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33106, "SRR29654140", "SRX25158192", "SRS21848814", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 2", "GSM8369973", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO", "GSM8369973", "GSM8369973: Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 2; Danio rerio; RNA Seq", "GSM8369973 r1", "GSM8369973", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L004_8hpi_DMSO_-005_cbc.fastq.gz", "fastq", 294091920.0, 4901532.0, "GSM8369973 r4", "0:60", "A:113032051;C:53520496;G:54754695;T:72762244;N:22434", 60, null, null, null, 113032051, 53520496, 54754695, 72762244, 22434, "SRX25158192", "SRS21848814", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33107, "SRR29654141", "SRX25158191", "SRS21848813", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 1", "GSM8369972", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO", "GSM8369972", "GSM8369972: Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 1; Danio rerio; RNA Seq", "GSM8369972 r1", "GSM8369972", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L001_8hpi_DMSO_-004_cbc.fastq.gz", "fastq", 79542600.0, 1325710.0, "GSM8369972 r1", "0:60", "A:29052800;C:15106261;G:15344010;T:20023888;N:15641", 60, null, null, null, 29052800, 15106261, 15344010, 20023888, 15641, "SRX25158191", "SRS21848813", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33108, "SRR29654142", "SRX25158191", "SRS21848813", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 1", "GSM8369972", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO", "GSM8369972", "GSM8369972: Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 1; Danio rerio; RNA Seq", "GSM8369972 r1", "GSM8369972", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L002_8hpi_DMSO_-004_cbc.fastq.gz", "fastq", 78150840.0, 1302514.0, "GSM8369972 r2", "0:60", "A:28490299;C:14795977;G:15298357;T:19554765;N:11442", 60, null, null, null, 28490299, 14795977, 15298357, 19554765, 11442, "SRX25158191", "SRS21848813", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33109, "SRR29654143", "SRX25158191", "SRS21848813", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 1", "GSM8369972", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO", "GSM8369972", "GSM8369972: Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 1; Danio rerio; RNA Seq", "GSM8369972 r1", "GSM8369972", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L003_8hpi_DMSO_-004_cbc.fastq.gz", "fastq", 80719140.0, 1345319.0, "GSM8369972 r3", "0:60", "A:29548150;C:15350378;G:15486892;T:20325654;N:8066", 60, null, null, null, 29548150, 15350378, 15486892, 20325654, 8066, "SRX25158191", "SRS21848813", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33110, "SRR29654144", "SRX25158191", "SRS21848813", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 1", "GSM8369972", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:8 xxx post injury DMSO", "GSM8369972", "GSM8369972: Neutrophils mpx+  8 xxx post injury  DMSO  biol rep 1; Danio rerio; RNA Seq", "GSM8369972 r1", "GSM8369972", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L004_8hpi_DMSO_-004_cbc.fastq.gz", "fastq", 79070760.0, 1317846.0, "GSM8369972 r4", "0:60", "A:28874779;C:14986197;G:15407453;T:19795922;N:6409", 60, null, null, null, 28874779, 14986197, 15407453, 19795922, 6409, "SRX25158191", "SRS21848813", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33111, "SRR29654145", "SRX25158190", "SRS21848812", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  DMSO  biol rep 3", "GSM8369971", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  DMSO  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO", "GSM8369971", "GSM8369971: Neutrophils mpx+  uninjured  DMSO  biol rep 3; Danio rerio; RNA Seq", "GSM8369971 r1", "GSM8369971", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L001_Uninjured_DMSO_-009_cbc.fastq.gz", "fastq", 182391600.0, 3039860.0, "GSM8369971 r1", "0:60", "A:65842196;C:33929963;G:37226765;T:45358269;N:34407", 60, null, null, null, 65842196, 33929963, 37226765, 45358269, 34407, "SRX25158190", "SRS21848812", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33112, "SRR29654146", "SRX25158190", "SRS21848812", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  DMSO  biol rep 3", "GSM8369971", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  DMSO  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO", "GSM8369971", "GSM8369971: Neutrophils mpx+  uninjured  DMSO  biol rep 3; Danio rerio; RNA Seq", "GSM8369971 r1", "GSM8369971", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L002_Uninjured_DMSO_-009_cbc.fastq.gz", "fastq", 179081640.0, 2984694.0, "GSM8369971 r2", "0:60", "A:64544585;C:33163305;G:37078756;T:44266486;N:28508", 60, null, null, null, 64544585, 33163305, 37078756, 44266486, 28508, "SRX25158190", "SRS21848812", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33113, "SRR29654147", "SRX25158190", "SRS21848812", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  DMSO  biol rep 3", "GSM8369971", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  DMSO  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO", "GSM8369971", "GSM8369971: Neutrophils mpx+  uninjured  DMSO  biol rep 3; Danio rerio; RNA Seq", "GSM8369971 r1", "GSM8369971", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L003_Uninjured_DMSO_-009_cbc.fastq.gz", "fastq", 185068740.0, 3084479.0, "GSM8369971 r3", "0:60", "A:66919946;C:34450944;G:37652300;T:46025841;N:19709", 60, null, null, null, 66919946, 34450944, 37652300, 46025841, 19709, "SRX25158190", "SRS21848812", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33114, "SRR29654148", "SRX25158190", "SRS21848812", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  DMSO  biol rep 3", "GSM8369971", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  DMSO  biol rep 3", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO", "GSM8369971", "GSM8369971: Neutrophils mpx+  uninjured  DMSO  biol rep 3; Danio rerio; RNA Seq", "GSM8369971 r1", "GSM8369971", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L004_Uninjured_DMSO_-009_cbc.fastq.gz", "fastq", 181507740.0, 3025129.0, "GSM8369971 r4", "0:60", "A:65513514;C:33675079;G:37432606;T:44872485;N:14056", 60, null, null, null, 65513514, 33675079, 37432606, 44872485, 14056, "SRX25158190", "SRS21848812", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33115, "SRR29654149", "SRX25158189", "SRS21848811", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  DMSO  biol rep 2", "GSM8369970", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  DMSO  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO", "GSM8369970", "GSM8369970: Neutrophils mpx+  uninjured  DMSO  biol rep 2; Danio rerio; RNA Seq", "GSM8369970 r1", "GSM8369970", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L001_Uninjured_DMSO_-006_cbc.fastq.gz", "fastq", 45794760.0, 763246.0, "GSM8369970 r1", "0:60", "A:16928569;C:8492491;G:9006326;T:11358172;N:9202", 60, null, null, null, 16928569, 8492491, 9006326, 11358172, 9202, "SRX25158189", "SRS21848811", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33116, "SRR29654150", "SRX25158189", "SRS21848811", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  DMSO  biol rep 2", "GSM8369970", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  DMSO  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO", "GSM8369970", "GSM8369970: Neutrophils mpx+  uninjured  DMSO  biol rep 2; Danio rerio; RNA Seq", "GSM8369970 r1", "GSM8369970", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L002_Uninjured_DMSO_-006_cbc.fastq.gz", "fastq", 45009960.0, 750166.0, "GSM8369970 r2", "0:60", "A:16608407;C:8309766;G:8990622;T:11094414;N:6751", 60, null, null, null, 16608407, 8309766, 8990622, 11094414, 6751, "SRX25158189", "SRS21848811", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33117, "SRR29654151", "SRX25158189", "SRS21848811", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  DMSO  biol rep 2", "GSM8369970", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  DMSO  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO", "GSM8369970", "GSM8369970: Neutrophils mpx+  uninjured  DMSO  biol rep 2; Danio rerio; RNA Seq", "GSM8369970 r1", "GSM8369970", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L003_Uninjured_DMSO_-006_cbc.fastq.gz", "fastq", 46565880.0, 776098.0, "GSM8369970 r3", "0:60", "A:17217451;C:8640544;G:9146095;T:11557146;N:4644", 60, null, null, null, 17217451, 8640544, 9146095, 11557146, 4644, "SRX25158189", "SRS21848811", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33118, "SRR29654152", "SRX25158189", "SRS21848811", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  DMSO  biol rep 2", "GSM8369970", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  DMSO  biol rep 2", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO", "GSM8369970", "GSM8369970: Neutrophils mpx+  uninjured  DMSO  biol rep 2; Danio rerio; RNA Seq", "GSM8369970 r1", "GSM8369970", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L004_Uninjured_DMSO_-006_cbc.fastq.gz", "fastq", 45624660.0, 760411.0, "GSM8369970 r4", "0:60", "A:16852187;C:8431444;G:9090234;T:11246907;N:3888", 60, null, null, null, 16852187, 8431444, 9090234, 11246907, 3888, "SRX25158189", "SRS21848811", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33119, "SRR29654153", "SRX25158188", "SRS21848810", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  DMSO  biol rep 1", "GSM8369969", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  DMSO  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO", "GSM8369969", "GSM8369969: Neutrophils mpx+  uninjured  DMSO  biol rep 1; Danio rerio; RNA Seq", "GSM8369969 r1", "GSM8369969", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L001_Uninjured_DMSO_-003_cbc.fastq.gz", "fastq", 55867380.0, 931123.0, "GSM8369969 r1", "0:60", "A:20680346;C:10254613;G:10895370;T:14026951;N:10100", 60, null, null, null, 20680346, 10254613, 10895370, 14026951, 10100, "SRX25158188", "SRS21848810", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33120, "SRR29654154", "SRX25158188", "SRS21848810", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  DMSO  biol rep 1", "GSM8369969", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  DMSO  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO", "GSM8369969", "GSM8369969: Neutrophils mpx+  uninjured  DMSO  biol rep 1; Danio rerio; RNA Seq", "GSM8369969 r1", "GSM8369969", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L002_Uninjured_DMSO_-003_cbc.fastq.gz", "fastq", 54889080.0, 914818.0, "GSM8369969 r2", "0:60", "A:20264110;C:10026239;G:10861715;T:13728039;N:8977", 60, null, null, null, 20264110, 10026239, 10861715, 13728039, 8977, "SRX25158188", "SRS21848810", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33121, "SRR29654155", "SRX25158188", "SRS21848810", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  DMSO  biol rep 1", "GSM8369969", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  DMSO  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO", "GSM8369969", "GSM8369969: Neutrophils mpx+  uninjured  DMSO  biol rep 1; Danio rerio; RNA Seq", "GSM8369969 r1", "GSM8369969", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L003_Uninjured_DMSO_-003_cbc.fastq.gz", "fastq", 56764440.0, 946074.0, "GSM8369969 r3", "0:60", "A:21036899;C:10416984;G:11027830;T:14275499;N:7228", 60, null, null, null, 21036899, 10416984, 11027830, 14275499, 7228, "SRX25158188", "SRS21848810", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [33122, "SRR29654156", "SRX25158188", "SRS21848810", "SRP517033", "PRJNA1129880", "Neutrophil immune profile guides spinal cord regeneration in zebrafish", "GSE271113", "Transcriptome Analysis", "Spinal cord injury triggers a strong innate inflammatory response in both non regenerative mammals and regenerative zebrafish. Neutrophils are the first immune population to be recruited to the injury site. Yet  their role in the repair process  particularly in a regenerative context  remains largely unknown. Here  we show that  promoting neutrophil inflammation resolution by inhibiting Cxcr4 boosts cellular and functional regeneration. Neutrophil specific RNA seq analysis reveals an enhanced activation state that correlates with a transient increase in tnf a expression in macrophage/microglia populations. Conversely  blocking neutrophil recruitment through Cxcr1/2 inhibition diminishes the presence of macrophage/microglia at the injury site and impairs spinal cord regeneration. Altogether  these findings provide new insights into the role of neutrophils in spinal cord regeneration  emphasizing the significant impact of their immune profile on the outcome of the repair process. Overall design: To understand the biological mechanisms promoted by neutrophils that improve spinal cord regeneration upon Cxcr4 inhibition  we sorted mpx:GFP+ cells from the injury site of lesioned larvae and from the same region of age matched uninjured controls and performed Bulk RNA sequencing analysis in AMD3100  and DMSO treated samples. Grant ID: 2022.02766.PTDC Grant title: Improving Spinal Cord Regeneration by modulating Neutrophil inflammation: lessons from a Zebrafish perspective Funding agency: Funda\u00e7\u00e5o para a Ci\u00eancia e Tecnologia FCT  Portugal", null, "pubmed:38925414", null, "Neutrophils mpx+  uninjured  DMSO  biol rep 1", "GSM8369969", null, "tissue:Neutrophils mpx:GFP+|cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO|geo loc name:missing|collection date:missing", "Neutrophils mpx+  uninjured  DMSO  biol rep 1", "Libraries were paired end sequenced on a Nextseq 500 Illumina  high output  with a 1x75 bp Illumina Kit Read 1: 26 cycles  index read: 6 cycles  Read 2: 60 cycles. Read 1 was used to identify the Illumina library index and CEL Seq sample barcode and read 2 was aligned to the GRCz11 reference transcriptome using Burrow Wheeler aligner BWA MEM. Reads that mapped equally well to multiple locations were discarded.  Mapping and generation of count tables was done using the MapAndGo script. Normalization and differential gene expression analyses were conducted using DESeq2 v1.38 Assembly: GRCz11 zebrafish Supplementary files format and content: counts table generated using the MapAndGo script", "Neutrophils mpx:GFP+", "At 3 dpf  larvae were anesthetized in 0.5 mM tricaine. Subsequenclty  the spinal cord was fully transected at the level of the anal pore using the tip of a 30G needle. Next  larvae were randomly distributed into different conditions and incubated with 25 uM AMD3100 supplemented with 0.1% DMSO from 4 xxx post injury hpi until 8 hpi.", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", "Zebrafish embryos were raised at 28.5\u00baC in E3 media with Methylene blue", "cell type:Neutrophils mpx:GFP+|treatment:uninjured DMSO", "GSM8369969", "GSM8369969: Neutrophils mpx+  uninjured  DMSO  biol rep 1; Danio rerio; RNA Seq", "GSM8369969 r1", "GSM8369969", "1", "Trunks containing the lesion site and same region of age matched uninjured controls were dissociated in a solution containing 2.5 mg/ml collagenase IV  15 mM HEPES  25 mM D Glucose  2% goat serum in HBSS without xxx and calcium  incubated at 28.5\u00baC shaking at 300 rpm  and vigorously dissociated using a pipette. Next they were filtered and stained with DAPI 1ul/ml for 10 min at 4C. Single cell suspensions were used to sort GFP+ cells directly into TRIzol using a FACSAria III BD Biosciences. All Samples were processed in less than two hours. Bulk RNA Sequencing was performed at Single Cell Discoveries. Total RNA was extracted from 2500 4800 GFP+ cells using the standard TRIzol protocol. mRNA was processed following an adapted version of CEL seq. Samples were barcoded with CEL seq primers during the reverse transcription and pooled post second strand synthesis TruSeq small RNA primers Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "NextSeq 500", null, "SRP517033", null, null, "IMM-CT-b001_HW5F2BGXN_S6_L004_Uninjured_DMSO_-003_cbc.fastq.gz", "fastq", 55531080.0, 925518.0, "GSM8369969 r4", "0:60", "A:20538193;C:10150017;G:10949669;T:13888674;N:4527", 60, null, null, null, 20538193, 10150017, 10949669, 13888674, 4527, "SRX25158188", "SRS21848810", "SRA1913211", "Instituto de Medicina Molecular", "Instituto de Medicina Molecular", null, null, null, null, null, null, null, null, null, null, null, "B", null, "usable mapping rate", "illumina", "nextseq", "unknown", "random_priming", "trueseq", "sc", "single_cell_plate", "celseq", null, "Portugal", "2024-06-29", "Larval", "Larval", "Blood", "Hematopoietic System"], [38122, "SRR1555598", "SRX684706", "SRS687823", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0106", "GSM1483841", null, "tissue:single embryo", "Metazome ZF timecourse sample 0106", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:4240", "GSM1483841", "GSM1483841: Metazome ZF timecourse sample 0106; Danio rerio; RNA Seq", "GSM1483841", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483841", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0106.fastq.gz", "fastq", 3911705.0, 111763.0, "GSM1483841 r1", "0:35", "A:1100064;C:790487;G:836206;T:1169251;N:15697", 35, null, null, null, 1100064, 790487, 836206, 1169251, 15697, "SRX684706", "SRS687823", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.72414, null, 0.09461, null, 0.96725, null, 0.4942, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38123, "SRR1555597", "SRX684705", "SRS687821", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0105", "GSM1483840", null, "tissue:single embryo", "Metazome ZF timecourse sample 0105", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:4200", "GSM1483840", "GSM1483840: Metazome ZF timecourse sample 0105; Danio rerio; RNA Seq", "GSM1483840", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483840", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0105.fastq.gz", "fastq", 32877145.0, 939347.0, "GSM1483840 r1", "0:35", "A:9524894;C:6625944;G:6696548;T:9953576;N:76183", 35, null, null, null, 9524894, 6625944, 6696548, 9953576, 76183, "SRX684705", "SRS687821", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.74436, null, 0.07095, null, 0.88201, null, 0.47612, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38124, "SRR1555596", "SRX684704", "SRS687822", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0104", "GSM1483839", null, "tissue:single embryo", "Metazome ZF timecourse sample 0104", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:4160", "GSM1483839", "GSM1483839: Metazome ZF timecourse sample 0104; Danio rerio; RNA Seq", "GSM1483839", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483839", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0104.fastq.gz", "fastq", 83810020.0, 2394572.0, "GSM1483839 r1", "0:35", "A:24075610;C:16799998;G:16694424;T:25967423;N:272565", 35, null, null, null, 24075610, 16799998, 16694424, 25967423, 272565, "SRX684704", "SRS687822", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.7588, null, 0.07993, null, 0.84449, null, 0.49785, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38125, "SRR1555595", "SRX684703", "SRS687820", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0103", "GSM1483838", null, "tissue:single embryo", "Metazome ZF timecourse sample 0103", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:4120", "GSM1483838", "GSM1483838: Metazome ZF timecourse sample 0103; Danio rerio; RNA Seq", "GSM1483838", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483838", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0103.fastq.gz", "fastq", 98206150.0, 2805890.0, "GSM1483838 r1", "0:35", "A:28361916;C:19232789;G:19589483;T:30798968;N:222994", 35, null, null, null, 28361916, 19232789, 19589483, 30798968, 222994, "SRX684703", "SRS687820", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.75509, null, 0.13429, null, 0.81117, null, 0.49102, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38126, "SRR1555594", "SRX684702", "SRS687819", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0102", "GSM1483837", null, "tissue:single embryo", "Metazome ZF timecourse sample 0102", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:4080", "GSM1483837", "GSM1483837: Metazome ZF timecourse sample 0102; Danio rerio; RNA Seq", "GSM1483837", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483837", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0102.fastq.gz", "fastq", 192817835.0, 5509081.0, "GSM1483837 r1", "0:35", "A:55969077;C:36880901;G:37835921;T:61101558;N:1030378", 35, null, null, null, 55969077, 36880901, 37835921, 61101558, 1030378, "SRX684702", "SRS687819", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.73387, null, 0.17245, null, 0.80164, null, 0.48657, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38127, "SRR1555593", "SRX684701", "SRS687818", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0101", "GSM1483836", null, "tissue:single embryo", "Metazome ZF timecourse sample 0101", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:4040", "GSM1483836", "GSM1483836: Metazome ZF timecourse sample 0101; Danio rerio; RNA Seq", "GSM1483836", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483836", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0101.fastq.gz", "fastq", 374339455.0, 10695413.0, "GSM1483836 r1", "0:35", "A:112191846;C:73679996;G:73185024;T:114766825;N:515764", 35, null, null, null, 112191846, 73679996, 73185024, 114766825, 515764, "SRX684701", "SRS687818", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.83131, null, 0.16618, null, 0.77812, null, 0.49895, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38128, "SRR1555592", "SRX684700", "SRS687816", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0100", "GSM1483835", null, "tissue:single embryo", "Metazome ZF timecourse sample 0100", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:4000", "GSM1483835", "GSM1483835: Metazome ZF timecourse sample 0100; Danio rerio; RNA Seq", "GSM1483835", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483835", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0100.fastq.gz", "fastq", 15239420.0, 435412.0, "GSM1483835 r1", "0:35", "A:4361241;C:3088134;G:3269084;T:4498093;N:22868", 35, null, null, null, 4361241, 3088134, 3269084, 4498093, 22868, "SRX684700", "SRS687816", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.7702, null, 0.17879, null, 0.92904, null, 0.505, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38129, "SRR1555591", "SRX684699", "SRS687817", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0099", "GSM1483834", null, "tissue:single embryo", "Metazome ZF timecourse sample 0099", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3960", "GSM1483834", "GSM1483834: Metazome ZF timecourse sample 0099; Danio rerio; RNA Seq", "GSM1483834", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483834", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0099.fastq.gz", "fastq", 284196500.0, 8119900.0, "GSM1483834 r1", "0:35", "A:81991733;C:57440756;G:58362818;T:86010858;N:390335", 35, null, null, null, 81991733, 57440756, 58362818, 86010858, 390335, "SRX684699", "SRS687817", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.81284, null, 0.1715, null, 0.78468, null, 0.49472, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38130, "SRR1555590", "SRX684698", "SRS687815", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0098", "GSM1483833", null, "tissue:single embryo", "Metazome ZF timecourse sample 0098", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3920", "GSM1483833", "GSM1483833: Metazome ZF timecourse sample 0098; Danio rerio; RNA Seq", "GSM1483833", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483833", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0098.fastq.gz", "fastq", 244245015.0, 6978429.0, "GSM1483833 r1", "0:35", "A:70141725;C:48500061;G:50748992;T:74478239;N:375998", 35, null, null, null, 70141725, 48500061, 50748992, 74478239, 375998, "SRX684698", "SRS687815", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.79668, null, 0.17718, null, 0.79271, null, 0.45966, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38131, "SRR1555589", "SRX684697", "SRS687814", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0097", "GSM1483832", null, "tissue:single embryo", "Metazome ZF timecourse sample 0097", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3880", "GSM1483832", "GSM1483832: Metazome ZF timecourse sample 0097; Danio rerio; RNA Seq", "GSM1483832", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483832", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0097.fastq.gz", "fastq", 215347510.0, 6152786.0, "GSM1483832 r1", "0:35", "A:62928172;C:42420829;G:43902946;T:65803342;N:292221", 35, null, null, null, 62928172, 42420829, 43902946, 65803342, 292221, "SRX684697", "SRS687814", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.80605, null, 0.17396, null, 0.78873, null, 0.50241, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38132, "SRR1555588", "SRX684696", "SRS687813", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0096", "GSM1483831", null, "tissue:single embryo", "Metazome ZF timecourse sample 0096", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3840", "GSM1483831", "GSM1483831: Metazome ZF timecourse sample 0096; Danio rerio; RNA Seq", "GSM1483831", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483831", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0096.fastq.gz", "fastq", 13992230.0, 399778.0, "GSM1483831 r1", "0:35", "A:4077402;C:2837417;G:2856511;T:4200750;N:20150", 35, null, null, null, 4077402, 2837417, 2856511, 4200750, 20150, "SRX684696", "SRS687813", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.77543, null, 0.14778, null, 0.93288, null, 0.48967, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38133, "SRR1555587", "SRX684695", "SRS687811", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0095", "GSM1483830", null, "tissue:single embryo", "Metazome ZF timecourse sample 0095", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3800", "GSM1483830", "GSM1483830: Metazome ZF timecourse sample 0095; Danio rerio; RNA Seq", "GSM1483830", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483830", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0095.fastq.gz", "fastq", 290151435.0, 8290041.0, "GSM1483830 r1", "0:35", "A:86694666;C:56953853;G:57315219;T:88811589;N:376108", 35, null, null, null, 86694666, 56953853, 57315219, 88811589, 376108, "SRX684695", "SRS687811", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.79098, null, 0.18155, null, 0.79034, null, 0.51338, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38134, "SRR1555586", "SRX684694", "SRS687810", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0094", "GSM1483829", null, "tissue:single embryo", "Metazome ZF timecourse sample 0094", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3760", "GSM1483829", "GSM1483829: Metazome ZF timecourse sample 0094; Danio rerio; RNA Seq", "GSM1483829", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483829", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0094.fastq.gz", "fastq", 645422925.0, 18440655.0, "GSM1483829 r1", "0:35", "A:186408265;C:127218686;G:131406509;T:199399112;N:990353", 35, null, null, null, 186408265, 127218686, 131406509, 199399112, 990353, "SRX684694", "SRS687810", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.81184, null, 0.15572, null, 0.78496, null, 0.48733, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38135, "SRR1555585", "SRX684693", "SRS687812", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0093", "GSM1483828", null, "tissue:single embryo", "Metazome ZF timecourse sample 0093", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3720", "GSM1483828", "GSM1483828: Metazome ZF timecourse sample 0093; Danio rerio; RNA Seq", "GSM1483828", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483828", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0093.fastq.gz", "fastq", 82574205.0, 2359263.0, "GSM1483828 r1", "0:35", "A:24239984;C:16625180;G:16309055;T:25123568;N:276418", 35, null, null, null, 24239984, 16625180, 16309055, 25123568, 276418, "SRX684693", "SRS687812", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.75557, null, 0.14554, null, 0.82205, null, 0.49736, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38136, "SRR1555584", "SRX684692", "SRS687809", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0092", "GSM1483827", null, "tissue:single embryo", "Metazome ZF timecourse sample 0092", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3680", "GSM1483827", "GSM1483827: Metazome ZF timecourse sample 0092; Danio rerio; RNA Seq", "GSM1483827", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483827", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0092.fastq.gz", "fastq", 72186905.0, 2062483.0, "GSM1483827 r1", "0:35", "A:21349438;C:14361391;G:14472348;T:21766197;N:237531", 35, null, null, null, 21349438, 14361391, 14472348, 21766197, 237531, "SRX684692", "SRS687809", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.74348, null, 0.13614, null, 0.8309, null, 0.51328, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38137, "SRR1555583", "SRX684691", "SRS687808", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0091", "GSM1483826", null, "tissue:single embryo", "Metazome ZF timecourse sample 0091", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3640", "GSM1483826", "GSM1483826: Metazome ZF timecourse sample 0091; Danio rerio; RNA Seq", "GSM1483826", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483826", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0091.fastq.gz", "fastq", 18405940.0, 525884.0, "GSM1483826 r1", "0:35", "A:5374937;C:3766785;G:3774551;T:5451369;N:38298", 35, null, null, null, 5374937, 3766785, 3774551, 5451369, 38298, "SRX684691", "SRS687808", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.74005, null, 0.11089, null, 0.91494, null, 0.49823, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38138, "SRR1555582", "SRX684690", "SRS687807", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0090", "GSM1483825", null, "tissue:single embryo", "Metazome ZF timecourse sample 0090", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3600", "GSM1483825", "GSM1483825: Metazome ZF timecourse sample 0090; Danio rerio; RNA Seq", "GSM1483825", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483825", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0090.fastq.gz", "fastq", 141938265.0, 4055379.0, "GSM1483825 r1", "0:35", "A:41371650;C:28115945;G:29356068;T:42485478;N:609124", 35, null, null, null, 41371650, 28115945, 29356068, 42485478, 609124, "SRX684690", "SRS687807", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.71151, null, 0.17711, null, 0.82789, null, 0.52369, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38139, "SRR1555581", "SRX684689", "SRS687806", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0089", "GSM1483824", null, "tissue:single embryo", "Metazome ZF timecourse sample 0089", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3560", "GSM1483824", "GSM1483824: Metazome ZF timecourse sample 0089; Danio rerio; RNA Seq", "GSM1483824", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483824", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0089.fastq.gz", "fastq", 99569015.0, 2844829.0, "GSM1483824 r1", "0:35", "A:28763571;C:20065085;G:19332839;T:31088193;N:319327", 35, null, null, null, 28763571, 20065085, 19332839, 31088193, 319327, "SRX684689", "SRS687806", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.74177, null, 0.13103, null, 0.81854, null, 0.51184, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38140, "SRR1555580", "SRX684688", "SRS687805", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0088", "GSM1483823", null, "tissue:single embryo", "Metazome ZF timecourse sample 0088", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3520", "GSM1483823", "GSM1483823: Metazome ZF timecourse sample 0088; Danio rerio; RNA Seq", "GSM1483823", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483823", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0088.fastq.gz", "fastq", 168790055.0, 4822573.0, "GSM1483823 r1", "0:35", "A:49034449;C:32785565;G:33439918;T:52997820;N:532303", 35, null, null, null, 49034449, 32785565, 33439918, 52997820, 532303, "SRX684688", "SRS687805", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.73454, null, 0.15775, null, 0.8047, null, 0.50587, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38141, "SRR1555579", "SRX684687", "SRS687803", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0087", "GSM1483822", null, "tissue:single embryo", "Metazome ZF timecourse sample 0087", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3480", "GSM1483822", "GSM1483822: Metazome ZF timecourse sample 0087; Danio rerio; RNA Seq", "GSM1483822", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483822", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0087.fastq.gz", "fastq", 83155975.0, 2375885.0, "GSM1483822 r1", "0:35", "A:23985678;C:16871573;G:17048421;T:25086100;N:164203", 35, null, null, null, 23985678, 16871573, 17048421, 25086100, 164203, "SRX684687", "SRS687803", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.75055, null, 0.10258, null, 0.83952, null, 0.49914, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38142, "SRR1555578", "SRX684686", "SRS687804", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0086", "GSM1483821", null, "tissue:single embryo", "Metazome ZF timecourse sample 0086", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3440", "GSM1483821", "GSM1483821: Metazome ZF timecourse sample 0086; Danio rerio; RNA Seq", "GSM1483821", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483821", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0086.fastq.gz", "fastq", 36772155.0, 1050633.0, "GSM1483821 r1", "0:35", "A:10536765;C:7377251;G:7701105;T:11014088;N:142946", 35, null, null, null, 10536765, 7377251, 7701105, 11014088, 142946, "SRX684686", "SRS687804", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.745, null, 0.08312, null, 0.88209, null, 0.52632, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38143, "SRR1555577", "SRX684685", "SRS687802", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0085", "GSM1483820", null, "tissue:single embryo", "Metazome ZF timecourse sample 0085", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3400", "GSM1483820", "GSM1483820: Metazome ZF timecourse sample 0085; Danio rerio; RNA Seq", "GSM1483820", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483820", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0085.fastq.gz", "fastq", 142281650.0, 4065190.0, "GSM1483820 r1", "0:35", "A:41238155;C:28228882;G:28727190;T:43901346;N:186077", 35, null, null, null, 41238155, 28228882, 28727190, 43901346, 186077, "SRX684685", "SRS687802", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.80695, null, 0.14045, null, 0.80361, null, 0.5, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38144, "SRR1555576", "SRX684684", "SRS687800", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0084", "GSM1483819", null, "tissue:single embryo", "Metazome ZF timecourse sample 0084", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3360", "GSM1483819", "GSM1483819: Metazome ZF timecourse sample 0084; Danio rerio; RNA Seq", "GSM1483819", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483819", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0084.fastq.gz", "fastq", 147589295.0, 4216837.0, "GSM1483819 r1", "0:35", "A:42100013;C:30125860;G:31767504;T:43391728;N:204190", 35, null, null, null, 42100013, 30125860, 31767504, 43391728, 204190, "SRX684684", "SRS687800", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.80373, null, 0.08661, null, 0.83157, null, 0.51923, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38145, "SRR1555575", "SRX684683", "SRS687801", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0083", "GSM1483818", null, "tissue:single embryo", "Metazome ZF timecourse sample 0083", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3320", "GSM1483818", "GSM1483818: Metazome ZF timecourse sample 0083; Danio rerio; RNA Seq", "GSM1483818", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483818", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0083.fastq.gz", "fastq", 147243985.0, 4206971.0, "GSM1483818 r1", "0:35", "A:42533757;C:29874233;G:30500611;T:44132167;N:203217", 35, null, null, null, 42533757, 29874233, 30500611, 44132167, 203217, "SRX684683", "SRS687801", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.80738, null, 0.08642, null, 0.82219, null, 0.49396, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38146, "SRR1555574", "SRX684682", "SRS687799", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0082", "GSM1483817", null, "tissue:single embryo", "Metazome ZF timecourse sample 0082", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3280", "GSM1483817", "GSM1483817: Metazome ZF timecourse sample 0082; Danio rerio; RNA Seq", "GSM1483817", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483817", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0082.fastq.gz", "fastq", 96747735.0, 2764221.0, "GSM1483817 r1", "0:35", "A:26768730;C:19578598;G:21713242;T:28549716;N:137449", 35, null, null, null, 26768730, 19578598, 21713242, 28549716, 137449, "SRX684682", "SRS687799", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.80738, null, 0.09417, null, 0.85167, null, 0.50713, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38147, "SRR1555573", "SRX684681", "SRS687798", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0081", "GSM1483816", null, "tissue:single embryo", "Metazome ZF timecourse sample 0081", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3240", "GSM1483816", "GSM1483816: Metazome ZF timecourse sample 0081; Danio rerio; RNA Seq", "GSM1483816", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483816", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0081.fastq.gz", "fastq", 158402895.0, 4525797.0, "GSM1483816 r1", "0:35", "A:46939694;C:31610300;G:32050592;T:47571349;N:230960", 35, null, null, null, 46939694, 31610300, 32050592, 47571349, 230960, "SRX684681", "SRS687798", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.78709, null, 0.11916, null, 0.82351, null, 0.48714, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38148, "SRR1555572", "SRX684680", "SRS687797", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0080", "GSM1483815", null, "tissue:single embryo", "Metazome ZF timecourse sample 0080", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3200", "GSM1483815", "GSM1483815: Metazome ZF timecourse sample 0080; Danio rerio; RNA Seq", "GSM1483815", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483815", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0080.fastq.gz", "fastq", 1432375.0, 40925.0, "GSM1483815 r1", "0:35", "A:422951;C:297837;G:308257;T:400778;N:2552", 35, null, null, null, 422951, 297837, 308257, 400778, 2552, "SRX684680", "SRS687797", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.7103, null, 0.12471, null, 0.9811, null, 0.57619, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38149, "SRR1555571", "SRX684679", "SRS687796", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0079", "GSM1483814", null, "tissue:single embryo", "Metazome ZF timecourse sample 0079", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3160", "GSM1483814", "GSM1483814: Metazome ZF timecourse sample 0079; Danio rerio; RNA Seq", "GSM1483814", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483814", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0079.fastq.gz", "fastq", 171937640.0, 4912504.0, "GSM1483814 r1", "0:35", "A:49235964;C:34067309;G:35846095;T:52553397;N:234875", 35, null, null, null, 49235964, 34067309, 35846095, 52553397, 234875, "SRX684679", "SRS687796", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.79389, null, 0.14252, null, 0.81107, null, 0.50474, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38150, "SRR1555570", "SRX684678", "SRS687795", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0078", "GSM1483813", null, "tissue:single embryo", "Metazome ZF timecourse sample 0078", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3120", "GSM1483813", "GSM1483813: Metazome ZF timecourse sample 0078; Danio rerio; RNA Seq", "GSM1483813", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483813", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0078.fastq.gz", "fastq", 220246950.0, 6292770.0, "GSM1483813 r1", "0:35", "A:63723749;C:43666354;G:45504302;T:67009596;N:342949", 35, null, null, null, 63723749, 43666354, 45504302, 67009596, 342949, "SRX684678", "SRS687795", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.80813, null, 0.1512, null, 0.80174, null, 0.49937, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38151, "SRR1555569", "SRX684677", "SRS687794", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0077", "GSM1483812", null, "tissue:single embryo", "Metazome ZF timecourse sample 0077", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3080", "GSM1483812", "GSM1483812: Metazome ZF timecourse sample 0077; Danio rerio; RNA Seq", "GSM1483812", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483812", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0077.fastq.gz", "fastq", 93138675.0, 2661105.0, "GSM1483812 r1", "0:35", "A:27102533;C:18314712;G:18498218;T:29006764;N:216448", 35, null, null, null, 27102533, 18314712, 18498218, 29006764, 216448, "SRX684677", "SRS687794", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.75818, null, 0.11499, null, 0.81572, null, 0.50169, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38152, "SRR1555568", "SRX684676", "SRS687793", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0076", "GSM1483811", null, "tissue:single embryo", "Metazome ZF timecourse sample 0076", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3040", "GSM1483811", "GSM1483811: Metazome ZF timecourse sample 0076; Danio rerio; RNA Seq", "GSM1483811", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483811", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0076.fastq.gz", "fastq", 118512415.0, 3386069.0, "GSM1483811 r1", "0:35", "A:33806370;C:23395934;G:24547522;T:36374942;N:387647", 35, null, null, null, 33806370, 23395934, 24547522, 36374942, 387647, "SRX684676", "SRS687793", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.75326, null, 0.1088, null, 0.81667, null, 0.49341, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38153, "SRR1555567", "SRX684675", "SRS687792", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0075", "GSM1483810", null, "tissue:single embryo", "Metazome ZF timecourse sample 0075", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:3000", "GSM1483810", "GSM1483810: Metazome ZF timecourse sample 0075; Danio rerio; RNA Seq", "GSM1483810", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483810", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0075.fastq.gz", "fastq", 2520525.0, 72015.0, "GSM1483810 r1", "0:35", "A:818327;C:420638;G:441967;T:833315;N:6278", 35, null, null, null, 818327, 420638, 441967, 833315, 6278, "SRX684675", "SRS687792", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.7119, null, 0.22445, null, 0.97368, null, 0.51561, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38154, "SRR1555566", "SRX684674", "SRS687791", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0074", "GSM1483809", null, "tissue:single embryo", "Metazome ZF timecourse sample 0074", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:2960", "GSM1483809", "GSM1483809: Metazome ZF timecourse sample 0074; Danio rerio; RNA Seq", "GSM1483809", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483809", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0074.fastq.gz", "fastq", 95385710.0, 2725306.0, "GSM1483809 r1", "0:35", "A:27032549;C:18681500;G:19490852;T:29708978;N:471831", 35, null, null, null, 27032549, 18681500, 19490852, 29708978, 471831, "SRX684674", "SRS687791", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.75216, null, 0.09004, null, 0.83469, null, 0.48768, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38155, "SRR1555565", "SRX684673", "SRS687790", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0073", "GSM1483808", null, "tissue:single embryo", "Metazome ZF timecourse sample 0073", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:2920", "GSM1483808", "GSM1483808: Metazome ZF timecourse sample 0073; Danio rerio; RNA Seq", "GSM1483808", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483808", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0073.fastq.gz", "fastq", 149165835.0, 4261881.0, "GSM1483808 r1", "0:35", "A:42991931;C:29913953;G:30117935;T:45803552;N:338464", 35, null, null, null, 42991931, 29913953, 30117935, 45803552, 338464, "SRX684673", "SRS687790", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.76488, null, 0.10545, null, 0.80586, null, 0.5046, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38156, "SRR1555564", "SRX684672", "SRS687789", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0072", "GSM1483807", null, "tissue:single embryo", "Metazome ZF timecourse sample 0072", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:2880", "GSM1483807", "GSM1483807: Metazome ZF timecourse sample 0072; Danio rerio; RNA Seq", "GSM1483807", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483807", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0072.fastq.gz", "fastq", 203829675.0, 5823705.0, "GSM1483807 r1", "0:35", "A:61171233;C:40047517;G:44515900;T:57461403;N:633622", 35, null, null, null, 61171233, 40047517, 44515900, 57461403, 633622, "SRX684672", "SRS687789", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.65225, null, 0.10254, null, 0.82014, null, 0.50437, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38157, "SRR1555563", "SRX684671", "SRS687788", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0071", "GSM1483806", null, "tissue:single embryo", "Metazome ZF timecourse sample 0071", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:2840", "GSM1483806", "GSM1483806: Metazome ZF timecourse sample 0071; Danio rerio; RNA Seq", "GSM1483806", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483806", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0071.fastq.gz", "fastq", 107894360.0, 3082696.0, "GSM1483806 r1", "0:35", "A:30475809;C:22092085;G:22685204;T:32400616;N:240646", 35, null, null, null, 30475809, 22092085, 22685204, 32400616, 240646, "SRX684671", "SRS687788", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.77637, null, 0.10375, null, 0.81574, null, 0.50918, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38158, "SRR1555562", "SRX684670", "SRS687787", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0070", "GSM1483805", null, "tissue:single embryo", "Metazome ZF timecourse sample 0070", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:2800", "GSM1483805", "GSM1483805: Metazome ZF timecourse sample 0070; Danio rerio; RNA Seq", "GSM1483805", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483805", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0070.fastq.gz", "fastq", 102775820.0, 2936452.0, "GSM1483805 r1", "0:35", "A:29014067;C:20865233;G:21656956;T:30738746;N:500818", 35, null, null, null, 29014067, 20865233, 21656956, 30738746, 500818, "SRX684670", "SRS687787", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.7643, null, 0.09535, null, 0.8325, null, 0.51201, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38159, "SRR1555561", "SRX684669", "SRS687786", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0069", "GSM1483804", null, "tissue:single embryo", "Metazome ZF timecourse sample 0069", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:2760", "GSM1483804", "GSM1483804: Metazome ZF timecourse sample 0069; Danio rerio; RNA Seq", "GSM1483804", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483804", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0069.fastq.gz", "fastq", 178882445.0, 5110927.0, "GSM1483804 r1", "0:35", "A:51610621;C:35847602;G:36898756;T:54275519;N:249947", 35, null, null, null, 51610621, 35847602, 36898756, 54275519, 249947, "SRX684669", "SRS687786", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.81481, null, 0.1212, null, 0.80338, null, 0.48647, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38160, "SRR1555560", "SRX684668", "SRS687784", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0068", "GSM1483803", null, "tissue:single embryo", "Metazome ZF timecourse sample 0068", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:2720", "GSM1483803", "GSM1483803: Metazome ZF timecourse sample 0068; Danio rerio; RNA Seq", "GSM1483803", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483803", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0068.fastq.gz", "fastq", 289924775.0, 8283565.0, "GSM1483803 r1", "0:35", "A:85968202;C:57218562;G:58533597;T:87797178;N:407236", 35, null, null, null, 85968202, 57218562, 58533597, 87797178, 407236, "SRX684668", "SRS687784", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.78411, null, 0.12015, null, 0.80736, null, 0.5018, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"], [38161, "SRR1555559", "SRX684667", "SRS687785", "SRP045679", "PRJNA258656", "Danio rerio high resolution developmental transcriptomic time course", "GSE60619", "Transcriptome Analysis", "Classical embryological studies revealed that during mid embryogenesis vertebrates show similar morphologies. This \u201cphylotypic stage\u201d has recently received support from transcriptome analyses  which have also detected similar stages in nematodes and arthropods. A conserved stage in these three phyla has led us to ask if all animals pass through a universal definitive stage as a consequence of ancestral constraints on animal development. Previous work has suggested that HOX genes may comprise such a \u2018zootypic\u2019 stage  however this hypothetical stage has hitherto resisted systematic analysis. We have examined the embryonic development of ten different animals each of a fundamentally different phylum  including a segmented worm  a flatworm  a roundworm  a water bear  a fruitfly  a sea urchin  a zebrafish  a sea anemone  a sponge  and a comb jelly. For each species  we collected the embryonic transcriptomes at 100 different developmental stages and analyzed their gene expression profiles. We found dynamic gene expression across all of the species that is structured in a stage like manner. Strikingly  we found that animal embryology contains two dominant modules of zygotic expression in terms of their protein domain composition: one involving proliferation  and a second involving differentiation. The switch between these two modules involves induction of the zootype; which in addition to homeobox containing genes  also involves Wnt and Notch signaling as well as forkhead domain transcription factors. Our results provide a systematic characterization of animal universality and identify the points of embryological constraints and flexibility. Overall design: 106 single embryo samples", "parent bioproject:PRJNA287810", "pubmed:26886793", null, "Metazome ZF timecourse sample 0067", "GSM1483802", null, "tissue:single embryo", "Metazome ZF timecourse sample 0067", "Libraries were sequenced on the Illumina HiSeq2000 according to standard  paired end sequencing  protocols. Primary analysis done in RTA 1.17.20 1.13.48 Conversion from BCL to FASTQ and deumltiplexing using CASAVA 1.8 configureBclToFastq.pl  fastq cluster count 1234567890   mismatches 0   use bases mask Y15n I6n Y35n Filter and read trimming barcode minimum quality of 10. trimming of read2 to 35 bases   not required. CEL Seq Hashimshony  et a. 2012l demultiplexing of second mate  using first mate barcode  allowing no mismatches in barcode. bowtie2  version 2.1.0  against Zv9 genome Read counting with htseq count version 0.5.4p3. Using on Ensemble Zv9 annotations Genome build: Zv9 Supplementary files format and content: tabular Expression matrix", "single embryo", null, "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "zebrafish fertilization was performed in the lab of Karina Yaniv Weizmann Institute  Israel. Four female and 1 male Danio rerio fish were mixed in a breeder tank and encouraged to spawn by tilting the tank and using artificial trees. post females started to release their eggs males quickly released sperm and fertilized eggs were collected into Zebrafish embryo medium as described in REF. Fertilized eggs were sampled in a small volume of medium every 40 minutes from fertilization into the cap of an 1.5 ml Eppendorf tube. Excess water was removed using a micro mouthpipette and the embryo flash frozen in liquid nitrogen.", "time min post fertilization:2680", "GSM1483802", "GSM1483802: Metazome ZF timecourse sample 0067; Danio rerio; RNA Seq", "GSM1483802", null, "1", "RNA was isolated using TRIzol as previously described Levin et al.  2012. The CEL Seq protocol Hashimshony  et al. 2012 was used to amplify and sequence. CEL seq multiplexing barocdes were used.", "GEO Accession:GSM1483802", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP045679", null, null, "Metazome_ZF_timecourse_sample_0067.fastq.gz", "fastq", 12717355.0, 363353.0, "GSM1483802 r1", "0:35", "A:3757539;C:2504848;G:2608725;T:3830804;N:15439", 35, null, null, null, 3757539, 2504848, 2608725, 3830804, 15439, "SRX684667", "SRS687785", "SRA179995", "GEO", "Yanai, Biology, Technion - Israel Institute of Technology", 1, 0.77835, null, 0.12605, null, 0.92604, null, 0.45926, null, 35, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "sc", "single_cell_plate", "celseq", null, "Israel", "2014-08-21", "Zygote", "Embryo", "Embryo Imprecise", "All anatomical structures"]], "truncated": false, "filtered_table_rows_count": 289, "expanded_columns": [], "expandable_columns": [], "columns": ["rowid", "run.accession", "experiment.accession", 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