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The effect of HP1-gamma deficiency on primordial germ cells

ABSTRACT: We compared the gene expression profiles of the HP1 gamma -/- vs wild-type PGCs to examine the molecular mechanisms of HP1g in PGC development. Although HP1g is suggested to be involved in PGC proliferation, our microarray data indicated that HP1g does not directly control PGC cell-cycle progression by regulating cell cycle-related genes. However, many genes acting on differentiated tissues were up-regulated in the HP1g-/- PGCs. The mVenus-positive cells from the E11.5 genital ridges were isolated by flow cytometry (JSAN, Bay Bioscience). The total RNA was then prepared from the sorted primordial germ cells (about 500-3,500 cells per embryo) of wild-type or HP1?-/- embryos with an RNeasy micro kit (Qiagen). The RNA samples were pooled from 5 or 6 embryos of each genotype. Two independent pooled RNA samples of each genotypes were used to verify the reproducibility of the microarray analyses.

ORGANISM(S): Mus musculus

SUBMITTER: Masahide Asano

PROVIDER: E-GEOD-27294 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Loss of heterochromatin protein 1 gamma reduces the number of primordial germ cells via impaired cell cycle progression in mice.

Abe Kanae K Naruse Chie C Kato Tomoaki T Nishiuchi Takumi T Saitou Mitinori M Asano Masahide M

Biology of reproduction 20110720 5

Signals from extraembryonic tissues in mice determine which proximal epiblast cells become primordial germ cells (PGCs). After their specification, approximately 40 PGCs appear at the base of the allantoic bud and migrate to the genital ridges, where they expand to about 25 000 cells by Embryonic Day (E)13.5. The heterochromatin protein 1 (HP1) family members HP1alpha, HP1beta, and HP1gamma (CBX5, CBX1, and CBX3, respectively) are thought to induce heterochromatin structure and to regulate gene ...[more]

PMID: 21778144

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The effect of HP1-gamma deficiency on primordial germ cells

Project description:We compared the gene expression profiles of the HP1 gamma -/- vs wild-type PGCs to examine the molecular mechanisms of HP1g in PGC development. Although HP1g is suggested to be involved in PGC proliferation, our microarray data indicated that HP1g does not directly control PGC cell-cycle progression by regulating cell cycle-related genes. However, many genes acting on differentiated tissues were up-regulated in the HP1g-/- PGCs.

2011-08-01 | GSE27294 | GEO

Role of Ror2 in primordial germ cell migration

Project description:Primordial germ cells (PGCs), the embryonic precursors of eggs and sperm, are a unique model for identifying and studying regulatory mechanisms in singly migrating cells. From their time of specification to eventual colonization of the gonad, mouse PGCs traverse through and interact with many different cell types, including epithelial cells and mesenchymal tissues. Work in drosophila and zebrafish have identified many genes and signaling pathways involved in PGC migration, but little is known about this process in mammals. We have generated a point mutation in the Ror2 gene that we know disrupts primordial germ cell migration in the developing mouse embryo. We used microarray analysis to determine if this defect is mediated through genome-wide or pathway-specific transcriptional changes. We analyzed primordial germ cells (PGCs) from 4 wild-type (WT) and 4 Ror2Y324C/Y324C mutant embryos using Oct4-DPE-EGFP. PGCs were collected during their active migratory state at embryonic day 9.5 (somite range 20-25).

2014-08-08 | E-GEOD-60179 | biostudies-arrayexpress

PRC1 coordinates timing of sexual differentiation of female primordial germ cells

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 Affymetrix microarray. Primodial germ cells were purified from embryos carrying Oct4(deltaPE)-GFP transgene by FACS.

2013-03-14 | E-GEOD-42782 | biostudies-arrayexpress

Transcription profiling of mouse germ cells with conditional Nanog knockdown.

Project description:Single Cell Microarray <br>E10.5 genital ridges (TM at E7.5) incubated in 0.5mM EDTA/ PBS for 20 minutes at 37ï¾°C were transferred to 2% BSA/ PBS. PGCs were collected from the genital ridges pierced with fine glass needles. Released PGCs were identified by their morphological characteristics and transferred into lysis buffer with a mouth pipette. PGCs were genotyped with the remaining genital ridges. The amplified cDNA library of each PGC was classified by qPCR-based expression analyses of Nanog, Oct4, and Stella/Dppa3 (Kurimoto et al., 2007). cDNAs were labeled by in vitro transcription (Affymetrix). The cRNAs were hybridized with the GeneChip Mouse Genome 430 2.0 Array (Affymetrix). Data were analyzed using the Microsoft Excel and MeV (multiple experimental viewer) software. <br>

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2014-08-01 | E-GEOD-48897 | biostudies-arrayexpress

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2013-03-04 | E-GEOD-40412 | biostudies-arrayexpress

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Project description:Primordial germ cells (PGCs) are lineage restricted precursor cells of sperm and eggs. While avian PGCs from chicken can be cultured and modified to produce genome-edited chickens, the long-term culture of PGCs from other bird species has not been achieved. Here, we explored the effects of a cell-matrix interaction on the in vitro propagation of chicken PGCs. Blocking integrin signaling severely reduced the self-renewal of the PGC, indicating that a PGC-matrix interaction is an essential process for self-renewal. We investigated the properties of somatic cell differentiated PGCs and expression analyses suggested that the PGCs undergo a partial epithelial-to-mesenchymal transition caused by excess PGC-matrix interactions. Finally, we conducted a long-term culture of chicken PGCs with matrix components, resulting in significant induction of their somatic conversion. These results suggested that a high level of PGC-matrix interaction can cause somatic conversion, while a moderate level is essential for their proliferation. Overall, we identified a molecular aspect of self-renewal and maintaining undifferentiated states of avian PGCs.

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2024-05-31 | PXD050727 | Pride

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Project description:Xenopus primordial germ cells (PGCs) are determined by the presence of maternally derived germ plasm. Germ plasm components both protect PGCs from somatic differentiation and begin a unique gene expression program. Segregation of the germline from the endodermal lineage occurs during gastrulation and PGCs subsequently initiate zygotic transcription. However, the gene-networks that operate to both preserve the potential for totipotency and promote germline differentiation are poorly understood. Here, we utilized RNA-sequencing analysis to comprehensively interrogate PGC and neighboring endoderm cell RNAs after lineage segregation. We identified 1,865 transcripts enriched in PGCs compared to endoderm cells. Over 50% of maternal, vegetally-enriched transcripts were enriched in the PGC transcriptome, including sox7. PGC-directed sox7 knockdown and over-expression studies revealed an early requirement for sox7 in proper germ plasm localization, zygotic transcription, and PGC number. We identified oct60 as the most highly expressed and enriched OCT3/4 homologue in PGCs. Lastly, we compared the Xenopus PGC transcriptome with human PGC transcripts and showed that 80% of transcripts are conserved, identifying Xenopus as a relevant model system for understanding the gene-networks necessary for human germline development.

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