Effect Of FoxO3 On The MRNA Expression Of Genes Related With Apoptosis And Autophagy In Goose Follicular Granulosa Cells

FoxO3 gene is a member of Fox gene family.It plays a role of transcription factor in the body.It can be phosphorylated,acetylated,ubiquitinated and other ways of post-translational modification.It is widely distributed in the body,including heart,brain,spleen,kidney,testis,ovary,skeletal muscle,fat and other tissues.FoxO3 can participate in a variety of signal pathways,such as PI3 K / Akt and AMPK which play an important role in regulating apoptosis,autophagy,proliferation,differentiation and oxidative stress.So far,the role of FoxO3 in female animal reproduction is still limited to human,mouse,pig and other mammals,whilst the role of FoxO3 in female poultry reproductive regulation is still unknown.Our previous reports demonstrated that the expression of FoxO3 gene was different in the ovarian tissue of geese during the egg-laying period and ceased period.In order to understand the role of FoxO3 in female reproduction,the expression of FoxO3,apoptosis and autophagy related genes in the follicles of geese during different stages of egg-laying cycle was detected.Thereafter,the eukaryotic expression vector of FoxO3 gene was constructed and transfected into the granulosa cells.Subsequently,the effect of overexpressing FoxO3 gene on the expression of apoptosis and autophagy related genes was investigated.In this study,we first collected the ovarian tissues of the female geese during the pre-laying,early-laying and peak-laying period,separated the prehierarchical follicles,and detected the expression of FoxO3 and apoptosis and autophagy related genes by q RT-PCR.The primers were designed according to the FoxO3 gene sequence of anser deposited in NCBI,and the CDS sequence of FoxO3 gene was amplified.Following the pc DNA3.1-myc-foxo3 was constructed and transferred into the primary follicular granulosa cells of geese by transient transfection,the expression of apoptosis and autophagy related genes were detected by q RT-PCR.The results showed that the FoxO3 gene was expressed during pre-laying and egg-laying period,and the expression level was increased with the increase of follicle diameter during early-laying and peak-laying period.The apoptosis and autophagy related genes including Bcl2,Bax,caspase 3,Fas,Fas L,LC3,and Beclin 1 were expressed in different prehierarchical follicles.Among them,the expression level of caspase 3 and Bax was increased with the increase of follicle diameter,while the expression level of LC3 was decreased with the increase of follicle diameter.Furthermore,the expression of other genes was also changed dynamically with the increase of follicle diameter.After transfected with pc DNA3.1-myc-foxo3,the expression of Bax,BCL2,Caspase3 and Fas L in granulosa cells was increased significantly,the expression of LC3 and Beclin1 was decreased significantly.This study showed that the expression of FoxO3 and other apoptosis autophagy genes such as FoxO3,Caspase3,Bax,LC3 and Fas/Fasl showed dynamic changes during different egg-laying stages.The overexpression of FoxO3 could affect the m RNA expression of apoptosis and autophagy gene in goose primary follicular granulosa cells.This study will provide the foundation for further understanding the regulation of FoxO3 in follicular development and atresia of birds.