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雷帕霉素处理对牦牛孤雌生殖后卵母细胞体外成熟和胚胎发育的影响。

Effect of rapamycin treatment on oocyte in vitro maturation and embryonic development after parthenogenesis in yaks.

机构信息

College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, China.

College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, China; Gansu Province Livestock Embryo Engineering Research Center, China.

出版信息

Theriogenology. 2022 Nov;193:128-135. doi: 10.1016/j.theriogenology.2022.09.017. Epub 2022 Sep 18.

Abstract

Autophagy plays an important role in mammalian oocyte maturation and early embryonic development and rapamycin is well known for inducing autophagy. Although previous studies have reported the effects of rapamycin on oocytes in vitro maturation (IVM) in different species, few studies have been reported on the role of rapamycin in yak oocytes IVM and embryonic development. Therefore, the objective of this study was to examine the effect of rapamycin treatment on yak oocytes IVM and early embryonic development. Specifically, immature yak oocytes during IVM or parthenogenetic (PA) embryos were treated with different rapamycin concentrations to select an optimal dose. Then evaluated its effect on maturation rates, cleavage, and blastocyst formation rates, mitochondrial membrane potential, ROS levels. Related genes and proteins expression in matured oocytes and blastocysts were also evaluated. The results show that 10 nM rapamycin treatment during IVM significantly improved oocyte maturation rates of oocytes and blastocyst formation rates. Treatment with 10 nM rapamycin reduced ROS level but increased mitochondrial membrane potential. Correspondingly, mRNA and protein expressions of LC3, Beclin-1, and Bcl-2 up-regulated while Bax down-regulated in matured yak COCs. When parthenogenetic embryos were treated with different rapamycin concentrations, 10 nM rapamycin treatment showed higher 8-cell and blastocyst formation rates. Also, CDX2, POU5F1, SOX2, and Nanog levels in blastocysts were upregulated. In summary, our findings demonstrate that rapamycin treatment improves oocytes maturation probably by increasing mitochondrial membrane potential, reducing ROS levels, and regulating the apoptosis in mature yak oocytes. Rapamycin treatment also improves embryonic developmental competence in the yak.

摘要

自噬在哺乳动物卵母细胞成熟和早期胚胎发育中发挥重要作用,雷帕霉素是诱导自噬的知名物质。虽然先前的研究已经报道了雷帕霉素对不同物种卵母细胞体外成熟(IVM)的影响,但关于雷帕霉素在牦牛卵母细胞 IVM 和胚胎发育中的作用的研究较少。因此,本研究旨在探讨雷帕霉素处理对牦牛卵母细胞 IVM 和早期胚胎发育的影响。具体来说,在 IVM 或孤雌激活(PA)胚胎中,用不同浓度的雷帕霉素处理未成熟的牦牛卵母细胞,以选择最佳剂量。然后评估其对成熟率、分裂和囊胚形成率、线粒体膜电位、ROS 水平的影响。还评估了成熟卵母细胞和囊胚中相关基因和蛋白质的表达。结果表明,10 nM 雷帕霉素处理在 IVM 期间可显著提高卵母细胞成熟率和囊胚形成率。用 10 nM 雷帕霉素处理可降低 ROS 水平,但增加线粒体膜电位。相应地,成熟牦牛 COCs 中的 LC3、Beclin-1 和 Bcl-2 的 mRNA 和蛋白表达上调,而 Bax 下调。当用不同浓度的雷帕霉素处理孤雌激活胚胎时,10 nM 雷帕霉素处理显示出更高的 8 细胞和囊胚形成率。此外,囊胚中的 CDX2、POU5F1、SOX2 和 Nanog 水平上调。总之,我们的研究结果表明,雷帕霉素处理可通过增加线粒体膜电位、降低 ROS 水平和调节成熟牦牛卵母细胞中的凋亡来改善卵母细胞成熟。雷帕霉素处理还可提高牦牛胚胎的发育能力。

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