雌激素在C型钠肽介导的山羊卵母细胞成熟调控中的作用机制研究

雌激素和C型钠肽(CNP)在山羊卵母细胞成熟调控中具有重要作用。前期研究表明，低浓度或高浓度17β-雌二醇分别通过雌激素核受体(ER-α和ER-β)或雌激素膜受体(GPR30)对CNP维持山羊卵母细胞减数分裂阻滞发挥不同生理效应，其具体机制需要进一步解析。本项目针对雌激素核受体信号通路和膜受体信号通路调控CNP介导的山羊卵母细胞成熟的分子机制，以山羊卵巢、卵泡和卵母细胞为材料，应用免疫荧光、实时定量PCR、Western Blot、ELISA、电镜等技术，通过对卵巢、卵泡、卵丘细胞和卵母细胞雌激素核受体和膜受体及其信号通路分子、卵母细胞成熟和卵丘细胞扩展相关基因表达或磷酸化水平，CNP受体NPR2表达及其鸟苷酸环化酶活性，cAMP和cGMP水平，以及卵母细胞核成熟进程、胞质成熟标志及卵丘细胞扩展等方面的系统研究，从分子水平解释雌激素核受体/膜受体通路对CNP介导的山羊卵母细胞成熟调控机制。

Estrogen and C type natriuretic peptide (CNP) play an important role in regulating the maturation of goat oocytes. Previous studies have shown that low or high concentration of 17β-estradiol play different physiological effects on the maintenance of goat oocytes meiosis arrest mediated by CNP through nuclear estrogen receptor (ER-α and ER-β) or membrane estrogen receptor (GPR30) respectively. However, their specific mechanism needs further research. In this study, the molecular mechanism of CNP mediated maturation of goat oocytes in response to nuclear estrogen receptor signaling pathway and membrane estrogen receptor signaling pathway will be investigated. The goat ovary, follicle and oocyte are used as materials. By means of immunohistochemical/immunofluorescence, real-time quantitative PCR, Western Blot, ELISA, electron microscope and so on, we will systematicly study the expression or phosphorylation of nuclear estrogen receptor and membrane estrogen receptor and their signaling pathway related molecules, oocyte maturation related gene, cumulus cell expansion related gene and CNP receptor NPR2. Furthermore, the expression of NPR2, guanylate cyclase activity and cAMP and cGMP levels will be detected in follicle, cumulus cells and oocyte. On other hands, the process of nuclear maturation, cytoplasmic maturation markers and cumulus expansion condition will be analysised. Through these studies, we will explain the regulation mechanism of estrogen nuclear receptor/membrane receptor signaling pathway on goat oocytes maturation mediated by CNP at the molecular level.