College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu, China.
South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Research Institute of Fish Culture and Hydrobiology, Faculty of Fisheries and Protection of Waters, University of South Bohemia in České Budějovice, Vodňany, Czech Republic.
J Cell Physiol. 2019 Aug;234(10):17677-17689. doi: 10.1002/jcp.28394. Epub 2019 Feb 26.
The fusion of sperm and oocytes determines the fertilization competence and subsequent development of embryos, which, in turn, can be affected by various proteins and DNA methylation. However, several factors in this whole regulation process remain unknown, especially in yaks. Here, we report that fibroblast growth factor 10 (FGF10) is an important growth factor that can enhance the maturation rate of yak oocytes and the motility of frozen spermatozoa. Subsequent blastocyst quality was also improved by increasing the total cell number and level of pregnancy-associated protein in blastocysts. These effects were significantly high in the group that received the 5 ng/ml FGF10 treatment, during both in vitro maturation (IVM) and capacitation. Our data show that the effects of FGF10 were dose-dependent at vital steps of embryogenesis in vitro. Furthermore, quantitative polymerase chain reaction, western blot analysis, and immunofluorescence demonstrated that the levels of CD9, CD81, DNMT1, and DNMT3B in both mature cumulus-oocyte complexes and capacitated sperms were regulated by FGF10, which was also highly expressed in the group treated with 5 ng/ml FGF10 during both IVM and capacitation. From our present study, we concluded that FGF10 promotes yak oocyte fertilization competence and subsequent blastocyst quality, and could also regulate CD9, CD81, DNMT1, and DNMT3B to optimize sperm-oocyte interactions and DNA methylation during fertilization.
精子和卵子的融合决定了受精能力和随后胚胎的发育,而这些又可以受到各种蛋白质和 DNA 甲基化的影响。然而,整个调控过程中的几个因素仍然未知,尤其是在牦牛中。在这里,我们报告说成纤维细胞生长因子 10(FGF10)是一种重要的生长因子,可以提高牦牛卵母细胞的成熟率和冷冻精子的活力。通过增加囊胚中的总细胞数和妊娠相关蛋白水平,随后也改善了囊胚的质量。在体外成熟(IVM)和获能过程中,接受 5ng/ml FGF10 处理的组中,这些效果明显更高。我们的数据表明,FGF10 在体外胚胎发生的关键步骤中的作用呈剂量依赖性。此外,定量聚合酶链反应、western blot 分析和免疫荧光表明,FGF10 调节了成熟的卵丘-卵母细胞复合物和获能精子中的 CD9、CD81、DNMT1 和 DNMT3B 的水平,在 IVM 和获能过程中接受 5ng/ml FGF10 处理的组中,FGF10 的表达也很高。从我们目前的研究中,我们得出结论,FGF10 促进牦牛卵母细胞的受精能力和随后的囊胚质量,并且可以调节 CD9、CD81、DNMT1 和 DNMT3B,以优化受精过程中的精子-卵相互作用和 DNA 甲基化。