National Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou 510030, China.
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science, Guangzhou 510030, China.
Molecules. 2022 Oct 12;27(20):6830. doi: 10.3390/molecules27206830.
The quality of in vitro matured oocytes is inferior to that of in vivo matured oocytes, which translates to low developmental capacity of embryos derived from in vitro matured oocytes. The developmental potential of in vitro matured oocytes is usually impaired due to oxidative stress. Stromal cell-derived factor-l (SDF1) can reduce oxidative stress and inhibit apoptosis. The aim of this study was to investigate the effects of SDF1 supplementation during pig oocyte in vitro maturation (IVM) on subsequent embryo development, and to explore the acting mechanisms of SDF1 in pig oocytes. We found that the IVM medium containing 20 ng/mL SDF1 improved the maturation rate of pig oocytes, as well as the cleavage rate and blastocyst rate of embryos generated by somatic cell nuclear transfer, in vitro fertilization, and parthenogenesis. Supplementation of 20 ng/mL SDF1 during IVM decreased the ROS level, increased the mitochondrial membrane potential, and altered the expression of apoptosis-related genes in the pig oocytes. The porcine oocyte transcriptomic data showed that SDF1 addition during IVM altered the expression of genes enriched in the purine metabolism and TNF signaling pathways. SDF1 supplementation during pig oocyte IVM also upregulated the mRNA and protein levels of YY1 and TET1, two critical factors for oocyte development. In conclusion, supplementation of SDF1 during pig oocyte IVM reduces oxidative stress, changes expression of genes involved in regulating apoptosis and oocyte growth, and enhances the ability of in vitro matured pig oocytes to support subsequent embryo development. Our findings provide a theoretical basis and a new method for improving the developmental potential of pig in vitro matured oocytes.
体外成熟的卵母细胞的质量不如体内成熟的卵母细胞，这导致了体外成熟卵母细胞衍生的胚胎发育能力低。体外成熟卵母细胞的发育潜能通常因氧化应激而受损。基质细胞衍生因子-1（SDF1）可以减轻氧化应激并抑制细胞凋亡。本研究旨在探讨 SDF1 在猪卵母细胞体外成熟（IVM）过程中的补充对随后胚胎发育的影响，并探讨 SDF1 在猪卵母细胞中的作用机制。我们发现，含有 20ng/mL SDF1 的 IVM 培养基提高了猪卵母细胞的成熟率，以及体细胞细胞核移植、体外受精和孤雌生殖产生的胚胎的分裂率和囊胚率。在 IVM 期间补充 20ng/mL SDF1 降低了 ROS 水平，增加了线粒体膜电位，并改变了猪卵母细胞中凋亡相关基因的表达。猪卵母细胞转录组数据表明，IVM 期间添加 SDF1 改变了富含嘌呤代谢和 TNF 信号通路的基因表达。在猪卵母细胞 IVM 期间补充 SDF1 还上调了 YY1 和 TET1 的 mRNA 和蛋白水平，这两种基因是卵母细胞发育的关键因素。总之，在猪卵母细胞 IVM 期间补充 SDF1 可减轻氧化应激，改变参与调节凋亡和卵母细胞生长的基因表达，并增强体外成熟猪卵母细胞支持随后胚胎发育的能力。我们的研究结果为提高猪体外成熟卵母细胞的发育潜能提供了理论依据和新方法。
