Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv Israel.
The Unit for Tmcr, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv Israel.
Cell Cycle. 2022 Apr;21(8):792-804. doi: 10.1080/15384101.2022.2031427. Epub 2022 Feb 1.
Fertilization triggers physiological degradation of maternal-mRNAs, which are then replaced by embryonic transcripts. Ample evidence suggests that Argonaut 2 (AGO2) is a possible post-fertilization regulator of maternal-mRNAs degradation; but its role in degradation of maternal-mRNAs during oocyte maturation remains obscure. Fyn, a member of the Src family kinases (SFKs), and an essential factor in oocyte maturation, was reported to inhibit AGO2 activity in oligodendrocytes. Our aim was to examine the role of Fyn and AGO2 in degradation of maternal-mRNAs during oocyte maturation by either suppressing their activity with SU6656 - an SFKs inhibitor; or by microinjecting DN-Fyn RNA for suppression of Fyn and BCl-137 for suppression of AGO2. Batches of fifteen mouse oocytes or embryos were analyzed by qPCR to measure the expression level of nine maternal-mRNAs that were selected for their known role in oocyte growth, maturation and early embryogenesis. We found that Fyn/SFKs are involved in maintaining the stability of at least four pre-transcribed mRNAs in oocytes at the germinal vesicle (GV) stage, whereas AGO2 had no role at this stage. During oocyte maturation, eight maternal-mRNAs were significantly degraded. Inhibition of AGO2 prevented the degreadation of at least five maternal-mRNAs, whereas inhibition of Fyn/SFK prevented degradation of at least five Fyn maternal-mRNAs and two SFKs maternal-mRNAs; pointing at their role in promoting the physiological degradation which occurs during oocyte maturation. Our findings imply the involvement of Fyn/SFKs in stabilization of maternal-mRNA at the GV stage and the involvement of Fyn, SFKs and AGO2 in degradation of maternal mRNAs during oocyte maturation.
受精触发母体 mRNA 的生理降解,然后被胚胎转录本取代。大量证据表明 Argonaut 2 (AGO2) 可能是母体 mRNA 降解的受精后调节因子;但它在卵母细胞成熟过程中对母体 mRNA 降解的作用仍不清楚。Fyn,Src 家族激酶 (SFKs) 的成员,也是卵母细胞成熟的必需因素,据报道可抑制少突胶质细胞中 AGO2 的活性。我们的目的是通过用 SU6656(SFKs 抑制剂)抑制其活性;或通过微注射 DN-Fyn RNA 抑制 Fyn 和 BCl-137 抑制 AGO2,来研究 Fyn 和 AGO2 在卵母细胞成熟过程中对母体 mRNA 降解的作用。用 qPCR 分析 15 个小鼠卵母细胞或胚胎的批次,以测量选择的 9 个母体 mRNA 的表达水平,这些母体 mRNA 因其在卵母细胞生长、成熟和早期胚胎发生中的已知作用而被选择。我们发现 Fyn/SFKs 参与维持卵母细胞中至少四个转录前 mRNA 的稳定性在生发泡 (GV) 阶段,而 AGO2 在这个阶段没有作用。在卵母细胞成熟过程中,8 个母体 mRNA 显著降解。AGO2 抑制防止至少 5 个母体 mRNA 的降解,而 Fyn/SFK 抑制防止至少 5 个 Fyn 母体 mRNA 和 2 个 SFKs 母体 mRNA 的降解;表明它们在促进卵母细胞成熟过程中发生的生理降解中起作用。我们的研究结果表明 Fyn/SFKs 参与 GV 阶段母体 mRNA 的稳定,Fyn、SFKs 和 AGO2 参与卵母细胞成熟过程中母体 mRNA 的降解。