牦牛卵泡发育过程中卵泡液差异蛋白质组学研究

Yaks (Bos grunniens) are the specific cattle in the highland and cold pastoral areas of the Qinghai-Tibetan plateau, and the reproduction of yak has obvious seasonal. Based on the yak oocyte in vitro development potential is difference between breeding season and non-breeding seasons, proteomics was used to study protein changes of follicular fluid in the different periods of follicular development, to establish a high abundance of two-dimensional gel electrophoresis profiles, and to analyze follicular fluid composition differences in before and after of oocyte maturation, different season, and different diameter follicles. The associated proteins of follicular development and oocyte maturation were discovered by comparative proteomic, and analyzed the protein function, then to reveal the mechanism of follicular development and the regulatory mechanism of follicular fluid on oocyte maturation in yak. This project would establish the proteomic technical system of follicular fluid, construct differences protein databases, and obtain functional proteins in different period of follicular development and oocyte maturation in yak. Thus, yak seasonal reproduction laws were clarified from protein levels, and to provides technical references and theoretical basis for improving reproductive efficiency and optimization oocyte in vitro culture systems in yak.

牦牛是青藏高原高寒牧区的特有牛种，繁殖有明显的季节性。在证实繁殖季节与非繁殖季节牦牛卵母细胞体外发育潜能不同的基础上，利用蛋白质组学手段研究牦牛卵泡发育过程中卵泡液中蛋白质的变化，建立高丰度的双向电泳图谱，探析卵母细胞成熟前后、不同繁殖季节、不同卵泡大小卵泡液组分差异，通过比较蛋白质组学筛查卵泡发育及卵母细胞成熟相关蛋白，对蛋白进行比对及功能分析，揭示牦牛卵泡发育机理及卵泡液对卵母细胞成熟的调控机制。本项目将建立牦牛卵泡液样品的蛋白质组学研究技术体系，构建差异蛋白质数据库，获得牦牛卵泡发育及卵母细胞成熟过程中的相关功能蛋白，从蛋白质水平了解牦牛季节性繁殖规律，为提高牦牛繁殖效率、完善牦牛卵母细胞体外培养体系提供技术参考与理论依据。

牦牛是青藏高原高寒牧区的特有牛种，繁殖有明显的季节性。在证实繁殖季节与非繁殖季节牦牛卵母细胞体外发育潜能不同的基础上，利用蛋白质组学手段研究牦牛卵泡发育过程中卵泡液中蛋白质的变化，建立高丰度的双向电泳图谱，探析不同繁殖季节、不同卵泡大小卵泡液组分差异，通过比较蛋白质组学筛查卵泡发育及卵母细胞成熟相关蛋白，对蛋白进行比对及功能分析，揭示牦牛卵泡发育机理及卵泡液对卵母细胞成熟的调控机制。项目开展了6个方面的研究，包括牦牛卵泡液与血浆差异蛋白质双向电泳方法的建立与优化、基于双向电泳（2-DE）技术的牦牛卵泡液（繁殖季节）差异蛋白质组学研究、基于同位素标记相对与绝对定量（iTRAQ）技术的牦牛卵泡液（繁殖季节与非繁殖季节）差异蛋白质组学研究、牦牛卵母细胞及体外受精胚胎早期发育基因差异表达的研究、牦牛线粒体基因组测序分析、牦牛PRDM16基因克隆及生物信息学与差异表达分析等，研究建立了牦牛卵泡液蛋白质组学研究技术体系，构建了卵泡液差异蛋白质数据库，获得了牦牛卵泡发育及卵母细胞成熟过程中的相关功能蛋白，从蛋白质水平了解了牦牛季节性繁殖规律，为提高牦牛繁殖效率、完善牦牛卵母细胞体外培养和胚胎体外生产体系提供了技术参考与理论依据。