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ATAC-seq of zebrafish primordial germ cells and somatic cells during early embryogenesis

ABSTRACT: Zebrafish Primordial Germ Cells (PGCs) were tracked in the Tg(Buc-GFP) line where the germ plasm protein Buc is fused to GFP. GFP-positive cells were isolated via FACS and RNA-seq was performed on polyadenylated transcripts for PGCs and somatic cells at 256-cell, high, dome, 10 somites and prim-5 stages. Two hundred cells were used for each biological replicate.

INSTRUMENT(S): NextSeq 550

ORGANISM(S): Danio rerio

SUBMITTER: Fabio D'Orazio M.

PROVIDER: E-MTAB-8741 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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RNA-seq of zebrafish primordial germ cells and somatic cells during early embryogenesis

Project description:Zebrafish Primordial Germ Cells (PGCs) were tracked in the Tg(Buc-GFP) line where the germ plasm protein Buc is fused to GFP. GFP-positive cells were isolated via FACS and RNA-seq was performed on polyadenylated transcripts for PGCs and somatic cells at 256-cell, high, dome, 10 somites and prim-5 stages. Two hundred cells were used for each biological replicate. RNA-seq was also performed on embryos in where Tdrd7 translation was inhibited via morpholino treatment.

2020-12-24 | E-MTAB-8707 | biostudies-arrayexpress

RNA-seq of zebrafish primordial germ cells and somatic cells during early embryogenesis

Project description:In this rescue experiment, embryos were injected with a Tdrd7-targeting morpholino to block translation of tdrd7 RNA and simultaneously provided with a Tdrd7 morpholino resistant RNA.

2020-12-24 | E-MTAB-9857 | biostudies-arrayexpress

Reduced representation bisulfite sequencing (RRBS) for primordial germ cells and somatic cells during zebrafish early embryogenesis

Project description:Zebrafish Primordial Germ Cells (PGCs) and somatic cells were isolated via FACS from Buc-GFP fish strain. Reduced representation bisulfite sequencing (RRBS) was performed on PGCs and somatic cells at high(6000-7000 cells per replicate), dome (7500 cells per replicate)and prim-5 stages(4500-5000 cells per replicate). Two replicates were collected for each cell type and stage.

2020-12-24 | E-MTAB-9858 | biostudies-arrayexpress

Expression data from ESC and in vitro derived somatic cells and germ cells

Project description:We performed gene expression profiling on in vitro derived PGCs, undifferentiated ESCs, and somatic cells from the EB to examine germ cell expression in ESC-derived cells All cells were collected using fluorescence activated cell sorting to isolate SSEA1+/cKit+ ESCs, SSEA1+/cKitbright PGCs, Oct4-gfp+/cKitbright PGCs, and SSEA1-/cKit- somatic cells and Oct4-gfp-/cKit- somatic cells.

2011-12-15 | E-GEOD-33121 | biostudies-arrayexpress

PRC1 coordinates timing of sexual differentiation of female primordial germ cells (RNA-Seq)

Project description:In mammals, sex differentiation of primordial germ cells (PGCs) is determined by extrinsic cues from the environment1. In female PGCs, expression of Stimulated by retinoic acid 8 (Stra8) and meiosis are induced in response to retinoic acid (RA) provided by the mesonephroi2-4. Given the widespread role of RA signaling during development8,9, the molecular mechanism specifying the competence of PGCs to timely express Stra8 and enter meiosis are unknown2,10. Here we identify gene dosage dependent roles in PGC development for Ring1 and Rnf2, two central components of the Polycomb Repressive Complex 1 (PRC1)11,13. Both paralogs are essential for PGC development between day 10.5 and 11.5 of gestation. Rnf2 is subsequently required in female PGCs for maintaining high levels of Oct4 and Nanog expression6, and for preventing premature induction of meiotic gene expression and entry into meiotic prophase. Chemical inhibition of RA signaling partially suppresses precocious Oct4 down-regulation and Stra8 activation in Rnf2-deficient female PGCs. Chromatin immunoprecipitation analyses show that Stra8 is a direct target of PRC1 and PRC2 in PGCs. These data demonstrate the importance of PRC1 gene dosage in PGC development and in coordinating the timing of sex differentiation of female PGCs by antagonizing extrinsic RA signaling. Gene expression of mouse primordial germ cells was analysed using RNAseq method. Primodial germ cells were purified from embryos carrying Oct4(-delta-PE)-GFP transgene by FACS.

2013-03-14 | E-GEOD-42852 | biostudies-arrayexpress

Single-cell RNA sequencing of lck-GFP transgenic zebrafish

Project description:The present study was conducted in the frame of the EU-funded Graphene Flagship project. The aim is to evaluate the impact of graphene oxide (GO) on the (innate) immune system using zebrafish as a model. We previously performed single-cell RNA-sequencing of germ-free zebrafish embryos exposed to GO plus the microbial metabolite butyrate (BA). Here, we performed a follow up experiment using germ-free lck-GFP transgenic fish in which the zebrafish were exposed to GO plus BA at 5 dpf. The embryos were then dissociated and subsequently sorted on lck and submitted for single-cell RNA-sequencing using 10x Genomics.

2022-07-23 | E-MTAB-11991 | biostudies-arrayexpress

ATAC-seq data in mouse and zebrafish limbs

Project description:Assay for Transposable Accessible Chromatin (ATAC) reveals a genome wide view of areas of open chromatin at very high resolution, which are often associated with regulatory activity. The ATAC-seq technology uses a Tn5 transposase loaded with nex-generation sequencing primers in order to simultaneously fragment areas of open chromatin and ligate adapters.

2015-01-06 | E-GEOD-61065 | biostudies-arrayexpress

PRMT5 protects genomic integrity during global DNA demethylation in primordial germ cells and preimplantation embryos

Project description:Primordial germ cells (PGCs) and preimplantation embryos undergo epigenetic reprogramming, which entails comprehensive erasure of DNA methylation. We found that PRMT5, an arginine methyltransferase, translocates from the cytoplasm to the nucleus during this process. Here we show that conditional loss of PRMT5 in early PGCs caused complete male and female sterility, which is preceded by upregulation of LINE1 and IAP transposons and DNA damage response. Similarly, loss of maternal-zygotic PRMT5 also leads to IAP upregulation and early embryonic lethality. We detected the PRMT5-catalysed repressive H2A/H4R3me2s modification on LINE1 and IAP in early wildtype PGCs, directly implicating this mark in the maintenance of transposon silencing during DNA hypomethylation. PRMT5 subsequently translocates back to the cytoplasm of PGCs to participate in the previously described PIWI-interacting RNA (piRNA) pathway that promotes transposon silencing via de novo DNA re-methylation. Thus, PRMT5 has a novel direct role in genome defense during preimplantation development and in PGCs at the time of global DNA demethylation PGCs mRNA profiles of embryonic day 11.5 dpc control (Blimp1Cre;Prmt5flox/+) and Prmt5 germ cell knockout (Blimp1Cre;Prmt5 flox/flox) were generated by deep sequencing (SOLiD next generation sequencing).

2014-09-09 | E-GEOD-61216 | biostudies-arrayexpress

PRMT5 protects genomic integrity during global DNA demethylation in primordial germ cells and preimplantation embryos

2014-11-14 | E-GEOD-60875 | biostudies-arrayexpress

Molecular sexual identity of chicken in vitro derived pre-gonadal primordial germ cells

Project description:In poultry, in vitro derived primordial germ cells (PGCs) represent an important tool for management of genetic resources. However, several studies have highlighted sexual differences exhibited by PGCs through in vitro steps, which may compromise their reproductive capacities. To understand this phenomenon, we compared the proteome of pregonadal chicken male (ZZ) and female (ZW) PGCs expanded in vitro by quantitative proteomic analysis using a GeLC-MS/MS strategy. The proteins found to be differentially abundant in chicken male and female PGCs indicated their early sexual identity. Many of the proteins up-accumulated in male PGCs were encoded by genes strongly enriched in the sexual chromosome Z. This suggests that the known lack of dosage compensation of the transcription of Z-linked genes between sexes persists at protein level in PGCs, and that this may be a key factor of their autonomous sex differentiation. Male and female PGCs up-accumulated protein sets were associated with differential biological processes, and contained proteins biologically relevant for male and female germ cell development respectively. This study presents first evidence on early predetermined sex specific cell fate of chicken PGCs that will help to understand their sexual physiological specificities and enable more precise sex-specific adaptation of in vitro culture conditions.

2021-11-03 | PXD022452 | Pride

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