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DNA Damage Responses in Mouse Primordial Germ Cells

ABSTRACT: DNA Damage Responses in Mouse Primordial Germ Cells

PROVIDER: PRJNA641697 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12082918.fastq.gz	Fastqsanger.gz
	SRR12082919.fastq.gz	Fastqsanger.gz
	SRR12082920.fastq.gz	Fastqsanger.gz
	SRR12082921.fastq.gz	Fastqsanger.gz
	SRR12082922.fastq.gz	Fastqsanger.gz

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DNA Damage Responses in Mouse Primordial Germ Cells [RNA-Seq]

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DNA Damage Responses in Mouse Primordial Germ Cells [ncRNA-Seq]

Project description:Germ cells specified during fetal development form the foundation of the mammalian germline. These primordial germ cells (PGCs) undergo rapid proliferation, yet the germline is highly refractory to mutation accumulation compared to somatic cells. Importantly, while the presence of endogenous or exogenous DNA damage has the potential to impact PGCs, there is little known about how these cells respond to stressors. To better understand the DNA damage response (DDR) in these cells, we exposed pregnant mice to ionizing radiation (IR) at specific gestational time points and assessed the DDR in PGCs. Our results show that PGCs prior to sex determination lack a G1 cell cycle checkpoint. Additionally, the response to IR-induced DNA damage differs between female and male PGCs post-sex determination. IR of female PGCs caused uncoupling of germ cell differentiation and meiotic initiation, while male PGCs exhibited repression of piRNA metabolism and transposon de-repression.

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DNA Damage Responses in Mouse Primordial Germ Cells [RNA-Seq]

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Primordial germ cell DNA demethylation and development require DNA translesion synthesis

Project description:Primordial germ cell DNA demethylation and development require DNA translesion synthesis

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MCM8 interacts with DDX5 to promote R-loop resolution in primordial germ cells

Project description:MCM8 has emerged as a core gene in reproductive aging and is crucial for meiotic homologous recombination repair. It also safeguards genome stability by coordinating the replication stress response during mitosis, but its function in mitotic germ cells remains elusive. Here we found that the number of primordial germ cells (PGCs) decreased dramatically after MCM8 deficiency due to proliferation defects. We further demonstrated that MCM8 interacted with two known helicases DDX5 and DHX9 in PGCs, and loss of MCM8 led to R-loop accumulation by reducing the retention of these helicases at R-loops, thus inducing genome instability. Cells expressing premature ovarian insufficiency-causative mutants of MCM8 with decreased interaction with DDX5 displayed increased R-loop levels. These results suggest that MCM8 interacts with R-loop-resolving factors to prevent DNA damage in PGCs, thus maintaining PGC proliferation and establishing the reproductive reserve. Our findings thus reveal a novel role for MCM8 in PGC development and improve our understanding of reproductive aging caused by genome instability in mitotic germ cells.

2024-05-31 | PXD050727 | Pride

Primordial germ cell DNA demethylation and development require DNA translesion synthesis

Project description:Mutations in DNA damage response (DDR) factors are associated with human infertility, which affects up to 15% of the population. It remains unclear if the role of DDR is solely in meiosis. One pathway implicated in human fertility is DNA translesion synthesis (TLS), which allows replication impediments to be bypassed. We find that TLS is essential for pre-meiotic germ cell development in the embryo. Loss of the central TLS component, REV1, significantly inhibits the induction of human PGC-like cells (hPGCLCs). This is recapitulated in mice, where deficiencies in TLS initiation (Rev1-/- or PcnaK164R/K164R) or extension (Rev7-/-) result in a >150-fold reduction in the number of primordial germ cells (PGCs) and complete sterility. In contrast, the absence of TLS does not impact the growth, function, or homeostasis of somatic tissues. Surprisingly, we find a complete failure in both DNA demethylation, a critical step in germline epigenetic reprogramming, and in activation of the germ cell transcriptional program. Our findings show that for normal fertility, DNA repair is required not only for meiotic recombination but for completion of DNA demethylation and the progression of PGC development.

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hnRNPL-orchestrated alternative splicing governs germ cell integrity and primordial follicle pool establishment

Project description:The primordial follicle pool established during the perinatal period, serves as the lifelong source of oocytes, with its depletion leading to premature ovarian failure (POF). Germ cell development from primordial germ cells (PGCs) to primordial follicle formation is tightly regulated by transcription and extensive post-transcriptional alternative splicing (AS). However, the dynamics and role of AS during PGC development and follicle formation remains poorly understood. Analysis of single-cell transcriptomes from E14.5 to P5 mouse ovaries revealed dynamic AS changes and key splicing factors involved in primordial follicle formation. Here, we employed a conditional knockout (cKO) model to demonstrate that hnRNPL loss in pre-meiotic germ cells disrupts homologous synapsis, induces extensive DNA damage, and results in abnormal diplotene-stage germ cells. Further analysis revealed that hnRNPL deficiency reduces transcript levels of genes essential for primordial follicle maintenance, causes aberrant splicing and protein expression of synapsis-related genes. These cumulative effects culminate in POF and infertility. Our findings explore AS dynamics and highlight hnRNPL as a key regulator of AS in primordial follicle pool formation and activation.

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Project description:DNA methylation reprogramming of primordial germ cells (PGCs) is an essential step that affects the activation and inactivation of certain genes, therefore having a direct impact on the transcriptome of an individual. In this study, we have described the methylome landscape of porcine PGCs, characterizing the genomic elements that resist methylation erasure.

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Genome-wide DNA methylation reprogramming in porcine primordial germ cells

Project description:Genome-wide DNA methylation reprogramming in porcine primordial germ cells

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