DNA methylation can be considered the most prominent in controlling the gene expression responsible for the balance between cell proliferation and cell death. In this study, we aimed to analyze the distinct contributions of Dnmt1 and Dnmt3a enzymes in oocyte maturation, survival, autophagy, reactive oxygen species (ROS) production, and compensation capacity of Dnmt3b and Dnmt3l enzymes in mouse oocytes. Following confirming the suppression of Dnmt1or Dnmt3a through siRNA application, the assessment involved immunofluorescence staining for Dnmts, 5mC, p62, and ROS levels. Cell death rates showed a noticeable increase while oocyte maturation rates exhibited significant reduction. Global DNA methylation showed a decline, concomitant with elevated p62 and ROS levels upon Dnmt1 or Dnmt3a knockdown. Remarkably, silencing of Dnmt1 led to an upsurge in Dnmt3a expression, whereas Dnmt3a knockdown triggered an increase in Dnmt1 levels. Furthermore, Dnmt3l expression exhibited a notable decrease after silencing of either Dnmt1 or Dnmt3a, while Dnmt3b levels remained comparable between control and siRNA-treated groups. Collectively, this study underscores the pivotal roles of Dnmt1 and Dnmt3a in orchestrating various facets of oocyte development, encompassing maturation, survival, autophagy, and ROS production. These findings offer valuable insights into the intricate regulatory network governed by DNA methylation machinery within the context of oocyte physiology.

中文翻译：

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揭示 DNA 甲基化机制的影响：Dnmt1 和 Dnmt3a 在协调卵母细胞发育和细胞稳态中的作用

DNA甲基化被认为是控制细胞增殖和细胞死亡之间平衡的基因表达中最重要的。在本研究中，我们旨在分析 Dnmt1 和 Dnmt3a 酶在小鼠卵母细胞成熟、存活、自噬、活性氧 (ROS) 产生以及 Dnmt3b 和 Dnmt3l 酶补偿能力中的独特贡献。在通过 siRNA 应用确认 Dnmt1 或 Dnmt3a 的抑制后，评估涉及 Dnmts、5mC、p62 和 ROS 水平的免疫荧光染色。细胞死亡率显着增加，而卵母细胞成熟率显着降低。在 Dnmt1 或 Dnmt3a 敲除后，整体 DNA 甲基化显示下降，同时 p62 和 ROS 水平升高。值得注意的是，Dnmt1 的沉默导致 Dnmt3a 表达激增，而 Dnmt3a 敲低则引发 Dnmt1 水平增加。此外，在沉默 Dnmt1 或 Dnmt3a 后，Dnmt3l 表达显着降低，而对照组和 siRNA 处理组之间的 Dnmt3b 水平保持相当。总的来说，这项研究强调了 Dnmt1 和 Dnmt3a 在协调卵母细胞发育的各个方面（包括成熟、存活、自噬和 ROS 产生）中的关键作用。这些发现为卵母细胞生理学背景下 DNA 甲基化机制控制的复杂调控网络提供了宝贵的见解。