Studies On The Mechanism Of Epidermal Growth Factor Relate Goat Granulosa Cells Apoptosis By Endoplasmic Reticulum Stress Pathways

The presence of millions of follicles on the ovaries of mammals, only1%of follicles willbe selected for further development and ovulation. While more than99%of follicles undergoa degenerative process known as atresia at various stages of development. Therefore, greatlylimits the use of excellent breeders. Varieties cultivated in focus, while the use of improvedvarieties has become an important issue to be solved in the current production. Technicalhormone superovulation and oocyte maturation in vitro techniques can get more than just partof the normal estrous oocytes, while the vast majority of oocytes and follicular atresia withboth the loss of resources caused by the loss of good dams. In recent years, assistedreproductive technology, especially as nuclear transfer, transgenic technology continues toimprove, how to get a lot of quality oocytes has restricted the promotion and application ofnew technology bottlenecks. Granulosa cell apoptosis is the main cause of follicular atresia.Follicular atresia is a leading cause of various clinical animal infertility in some extent. EGFhas an important role in promoting follicle development. At present, the mechanism of EGFinhibiting apoptosis of goat follicular granulosa cells by the endoplasmic reticulum stress isnot clear. By collecting goats and isolated and cultured ovarian granulosa cells, granulosacells by constructing the endoplasmic reticulum stress model. Research EGF regulation goatgranulosa cell apoptosis, providing a theoretical foundation for the clinical mass high-qualityfollicles.The ovaries from goats were obtained at local slaughterhouse, granulosa cells werecollected from isolated that healthy follicles of3-5mm in diameter. By serum withdrawalinduced spontaneous apoptosis. By adding tunicamycin induce endoplasmic reticulum stress;Results of annexin V-FITC/PI staining examination for apoptotic goat granulosa cells culturedin vitro; The expression of the Grp78, ATF4, ATF6, CHOP and Caspase-3mRNA wasexamined by real-time quantitative PCR. The results showed that, Serum-free group and5μg/mL TM-tread group significantly induced apoptosis of granulosa cells, and atime-dependent manner;10ng/mL EGF significantly inhibited that apoptosis of granulosacells; EGF significantly lowered Caspase-3mRNA expression levels under serum-free culturegroup and5μg/mL TM group cultured for24hr; Grp78and CHOP mRNA significantly increase under serum-free culture group cultured for24and48hr. Meanwhile, EGFsignificantly inhibit Grp78, ATF4, ATF6and CHOP mRNA under serum-free culture groupand TM group.Endoplasmic reticulum (ER) stress could involved in granulosa cell apoptosis duringgoat follicular atresia. EGF down-regulating the expression of ATF4, ATF6and CHOP mRNAlevels, which inhibit goat granulosa cells apoptosis by endoplasmic reticulum stress. Theresults provide a theoretical reference to reveal the mechanism of the clinical follicular atresiaand infertility treatment.