Duan Jiaxin, Xu Anli, Xie Junhao, Wang Linghao, Lv Qipin, Yang Jingxian, Zhao Ning, Li Bugao, Cao Guoqing
College of Animal Science, Shanxi Agricultural University, Jinzhong, 030801, China; Shanxi Key Laboratory of Animal Genetics Resource Utilization and Breeding, Jinzhong, 030801, China.
College of Animal Science, Shanxi Agricultural University, Jinzhong, 030801, China; Shanxi Key Laboratory of Animal Genetics Resource Utilization and Breeding, Jinzhong, 030801, China.
Theriogenology. 2025 Oct 15;246:117537. doi: 10.1016/j.theriogenology.2025.117537. Epub 2025 Jun 15.
The G protein-coupled receptor 30 (GPR30) exhibits specific expression in oocytes and plays a pivotal role in regulating mammalian oocyte maturation. However, the molecular mechanisms underlying GPR30-mediated porcine oocyte maturation remain poorly characterized. This study elucidated the regulatory function of GPR30 during in vitro maturation (IVM) of porcine oocytes and its potential mechanistic basis. Western blot levels of GPR30 showed that GPR30 protein content decreased with time during oocyte development; however, the addition of 17β-estradiol for 24 h of incubation significantly increased GPR30 in porcine oocytes. At the same time, fluorescent staining results showed that the 17β-estradiol group improved the first polar body (PB1) rate, the cumulus expansion index (CEI), and the mitochondria and endoplasmic reticulum normal distribution rate. G15 is a specific antagonist of GPR30. We found that the 17β-estradiol + G15 groups inhibited the above facilitation. In vitro experiments with porcine oocytes further showed that autophagy was improved by the 17β-estradiol group, and the promotion of 17β-estradiol and the maturation of the oocyte nucleoplasm were impeded by the use of the MEK/ERK-specific inhibitors Trametinib and FR 180204, respectively. Our findings indicated that GPR30 positively regulates the occurrence of autophagy in porcine oocytes and affects oocyte maturation and development via the MEK/ERK signaling pathway. In summary, the study provides a theoretical foundation for the further understanding of the MEK/ERK signaling pathway as a regulatory mechanism for mammalian oocyte maturation.
G蛋白偶联受体30（GPR30）在卵母细胞中呈现特异性表达，并在调节哺乳动物卵母细胞成熟过程中发挥关键作用。然而，GPR30介导猪卵母细胞成熟的分子机制仍不清楚。本研究阐明了GPR30在猪卵母细胞体外成熟（IVM）过程中的调节功能及其潜在的机制基础。GPR30的蛋白质印迹水平显示，在卵母细胞发育过程中，GPR30蛋白含量随时间下降；然而，添加17β-雌二醇孵育24小时可显著增加猪卵母细胞中的GPR30。同时，荧光染色结果表明，17β-雌二醇组提高了第一极体（PB1）率、卵丘扩展指数（CEI）以及线粒体和内质网的正常分布率。G15是GPR30的特异性拮抗剂。我们发现，17β-雌二醇 + G15组抑制了上述促进作用。对猪卵母细胞的体外实验进一步表明，17β-雌二醇组改善了自噬，而分别使用MEK/ERK特异性抑制剂曲美替尼和FR 180204则阻碍了17β-雌二醇的促进作用以及卵母细胞核质的成熟。我们的研究结果表明，GPR30正向调节猪卵母细胞自噬的发生，并通过MEK/ERK信号通路影响卵母细胞的成熟和发育。总之，该研究为进一步理解MEK/ERK信号通路作为哺乳动物卵母细胞成熟的调节机制提供了理论基础。
