褪黑素调控绵羊卵泡颗粒细胞褪黑素受体表达的蛋白组学

当前研究显示褪黑素对褪黑素受体在卵泡颗粒细胞中的表达有促进作用，而其作用机制及对卵母细胞成熟的影响机制还不清楚。本项目拟以绵羊卵泡颗粒细胞为研究对象，应用细胞分离和培养、分子生物学、iTRAQ蛋白质谱定量分析等实验方法，研究：①褪黑素促进绵羊卵泡颗粒细胞MR 表达的浓度和时间；②褪黑素调控绵羊卵泡颗粒细胞蛋白组学和细胞外分泌蛋白组学；③差异表达的外分泌蛋白因子对卵母细胞体外成熟的影响。进而探讨褪黑素调控颗粒细胞的基因和蛋白表达的关联机制，这将为揭示褪黑素在卵泡发育过程中的生理作用和为进一步开发和利用哺乳动物的繁殖潜能奠定基础。

The recent researchs show that melatonin has amplification effect on melatonin receptor expressing in the follicular granulosa cells. However, its mechanism of the action and the influence about oocyte maturation is not very clear. Whitin sheep ovarian follicles granulosa cells, by using cell isolation and culture, molecular biology, iTRAQ protein spectrum quantitative analysis, etc., the aims of this project are to research: (1)the time- and dose-dependent manners of melatonin promote MR mRNA expressing in granulosa cells; (2) differential proteomics of melatonin regulating the granulosa cells protein and granular cells exocrine protein;(3) the influences of differentially expressed exocrine protein factors on oocytes maturation in vitro.And then discusses related mechanisms of melatonin regulating granulosa cells of gene and protein expression. These will provide solid and scientific proof for revealing physiological role of melatonin in follicular development and for the further development and utilization of mammalian reproductive potential.