Study Of CREBZF/Zhangfei On Cell Apoptosis In Mouse Ovary Granulosa Cells

In mammals,the development of follicles is a complex process.During folliculogenesis,many kinds of hormone include gonadal gonadal and growth factors induce ovulation.However,the large number of follicles atresia,only minimal follicles ovulate and form a corpus luteum.Follicular atresia begin at granular cell apoptosis.Extensive research reveal that follicular development is regulated by members of the mammalian ER stress response-related molecular chaperones.But the molecular mechanisms between the ER stress response and follicular development still unclear.CREBZF/Zhangfei is one of basic region leucine zipper transcription factor which found in studing of HCF-1.CREBZF is expressed in various organs.According to most reports,CREBZF/Zhangfei play a important role in unfolded protein response(UPR)and the ER stress response.,We study the locatizaion,function,expression rule and regulating mechanism of CREBZF/Zhangfei in the development of follicles by using immunohistochemistry,lentivirus,western blot,RT-PCR,flow cytometry.The main results were as follows:(1)The expression and localization of CREBZF/Zhangfei was found in the mouse ovarian follicle.The expression of CREBZF/Zhangfei has regularity in the reproductive cycle.The western blot results indicate that the expression of CREBZF/Zhangfei was higher at estrus.In addition,the expression of CREBZF was higher when artificial inducing the development of follicle.(2)By using constructed recombinant lentiviral vectors,we successed to manual control CREBZF/Zhangfei expressed in mouse ovary granular cell.the efficincy can be 70%.(3)By manual control of CREBZF/Zhangfei expressed in mouse ovary granular cell,we found that the ovary granular cell’s apoptosis rate was change in vitro.Further study shows that Zhangfei knockdown significantly increased the protein expression of pERK and BCL-2 and decreased the protein expression of GRP78,CHOP,ERK,BAX and Caspase-3.On the contrary,over expression of Zhangfei increased the protein expression of CHOP,ERK,BAX and Caspase-3,but decreased the protein expression of BCL-2 and pERK.In summary,this experiment studied the locatization,function,expression rule and regulation mechanism of CREBZF/Zhangfei in the development of follicles.This research create new experimental data for studying the function of CREBZF/Zhangfei in follicles development.