High-throughput stage-specific transcriptomics provides an unbiased approach for understanding the process of cell development. Here, we report transcriptome analysis of primordial germ cell, female germline stem cell (FGSC), germinal vesicle and mature oocyte by performing RNA sequencing of freshly isolated cells in mice. As expected, these stages and gene-expression profiles are consistent with developmental timing. Analysis of genome-wide DNA methylation during female germline development was used for confirmation. By pathway analysis and blocking experiments, we demonstrate PI3K-AKT pathway is critical for FGSC maintenance. We also identify functional modules with hub genes and lncRNAs, which represent candidates for regulating FGSC self-renewal and differentiation. Remarkably, we note alternative splicing patterns change dramatically during female germline development, with the highest occurring in FGSCs. These findings are invaluable resource for dissecting the molecular pathways and processes into oogenesis and will be wider applications for other types of stem cell research.

中文翻译：

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通过低输入细胞的 RNA 测序和全基因组 DNA 甲基化分析揭示早期雌性生殖细胞的分子特征。


高通量阶段特异性转录组学为理解细胞发育过程提供了一种公正的方法。在这里，我们通过对小鼠中新鲜分离的细胞进行 RNA 测序，报告了原始生殖细胞、雌性生殖干细胞 (FGSC)、生殖囊泡和成熟卵母细胞的转录组分析。正如预期的那样，这些阶段和基因表达谱与发育时间一致。对雌性种系发育过程中全基因组 DNA 甲基化的分析用于确认。通过通路分析和阻断实验，我们证明 PI3K-AKT 通路对于 FGSC 的维持至关重要。我们还鉴定了具有 hub 基因和 lncRNA 的功能模块，它们代表了调节 FGSC 自我更新和分化的候选者。值得注意的是，我们注意到选择性剪接模式在雌性种系发育过程中发生了巨大变化，其中最高的发生在 FGSC 中。这些发现是剖析卵子发生的分子途径和过程的宝贵资源，并将更广泛地应用于其他类型的干细胞研究。