Stella prevents excessive de novo DNA methylation during mouse oogenesis

Oocyte acquires developmental competence during its maturation. This stage is accompanied with large-scale alteration in transcription, and series of genome-wide epigenetic reprogramming, including de novo establishment of DNA methylation. However, our understanding of mechanisms regulating this process is limited. To investigate the role of Stella (Dppa3) in de novo methylation during mouse oogenesis, here we measured DNA methylation by RRBS and expression profiles by RNA-seq in PGCs and oocytes at serveral development stages, including genotypes of both Stella (Dppa3) +/- and Stella -/-. Overall design: We measured DNA methylation by RRBS and expression profiles by RNA-seq in PGCs and oocytes at serveral development stages (E13.5, D5, D10, D20, MII), including genotypes of both Stella (Dppa3) +/- and Stella -/-. DNA methylation of sperm with both genotypes were also measured by RRBS for comparision.

卵母细胞在成熟过程中获得发育潜能。该阶段伴随转录组的大规模动态重塑，以及一系列全基因组表观遗传重编程事件，包括DNA甲基化的从头建立。然而，目前学界对调控该过程的分子机制仍缺乏深入认知。为阐明斯特拉（Stella, Dppa3）在小鼠卵子发生过程中DNA甲基化从头建立环节的调控作用，本研究采用简化代表性重亚硫酸盐测序（RRBS）检测DNA甲基化水平，通过RNA测序（RNA-seq）分析基因表达谱，对多个发育阶段（E13.5、D5、D10、D20、MII）的原始生殖细胞（PGCs）与卵母细胞进行了检测，样本涵盖斯特拉（Stella, Dppa3）杂合型（+/-）与敲除纯合型（-/-）两种基因型。此外，本研究还对两种基因型的精子样本开展RRBS检测以作为对照。实验设计概述：我们通过RRBS检测DNA甲基化水平、通过RNA-seq分析基因表达谱，对多个发育阶段（E13.5、D5、D10、D20、MII）的PGCs与卵母细胞进行了检测，样本涵盖斯特拉（Stella, Dppa3）+/-与-/-两种基因型；同时对两种基因型的精子样本进行RRBS检测以作对照。