Ghrelin对牛卵母细胞体外成熟的调控机制研究

项目名称： Ghrelin对牛卵母细胞体外成熟的调控机制研究

项目编号： No.31301975

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 许晓玲

作者单位： 北京市农林科学院

项目金额： 24万元

中文摘要： Ghrelin，又名生长激素释放肽，其参与了性腺轴中激素的分泌和调节，并对斑马鱼、猪和绵羊卵母细胞的体外成熟起着重要作用，但具体的作用机制还不清楚，本项目拟围绕Ghrelin在牛卵母细胞体外成熟中的作用及其调控机制进行研究。首先通过mRNA原位杂交以及免疫组化检测Ghrelin及其受体在卵巢中的表达和定位，其次通过添加不同浓度的Ghrelin来确定其对牛卵母细胞体外成熟的影响；通过添加Ghrelin和其受体拮抗剂D-lys3-GHRP-6共培养来确定Ghrelin对卵母细胞体外成熟的作用途径；并进一步研究Ghrelin对牛颗粒细胞和卵母细胞类固醇激素（E2和P4）分泌、cAMP水平以及对MPF和MAPK活性的影响，揭示其作用的分子机制。研究结果将对解释复杂的哺乳动物卵母细胞体外成熟过程提供重要理论依据，同时也为家畜胚胎体外生产提供有力的技术保障，因此本项目具有重要的学术价值和应用前景。

中文关键词： 牛；生长激素释放肽；卵母细胞；体外成熟；调控机制

英文摘要： Ghrelin, also known as growth hormone releasing peptide, which participates in the hormone secretion and the regulation of gonadal axis，and also plays an role in vitro maturation of zebrafish，porcine and ovis aries oocytes, but the exact mechanism of action is unclear. This project aims to study the role of Ghrelin in bovine oocyte maturation in vitro and its regulation mechanism. Firstly, mRNA in situ hybridization and immunohistochemistry are used to examine the expression and localization of Ghrelin and its receptor in ovary,followed by adding different concentrations of exogenous Ghrelin, the maturation rate of oocytes to determine its effects on bovine oocytes maturation in vitro;Ghrelin and its receptor antagonist D-lys3-GHRP-6 were added to determine the pathway of Ghrelin on oocyte; effects of Ghrelin on bovine granulosa cells and oocyte steroid hormone ( E2 and P4 ) secretion, cAMP levels, as well as the influence on MPF and MAPK activity, molecular mechanism of its function. The findings will provide important theoretical basis for further explanation of mammalian oocytes maturation in vitro, but also provide powerful technical support for the animal embryo in vitro production, so the project has important academic value and application prospect.

英文关键词： Bovine；Ghrelin；oocyte；in vitro maturation；regulatory mechanism