| Objective: The expression of Semaphorin6 C is decreasing with the development of ovarian aging, suggesting that Semaphorin6 C may play an important role in ovarian aging process, and the percentage of atresia follicles rate in older ovaries is exclusively high. In this study, we plan to examine whether semaphorin6 c has effects on cell junctions during follicular atresia, and try to explore the mechanism.Methods: 30 14-day old C57 mice as research object were selected and isolated preantral follicles through stereomicroscope using 3D-method. Preantral follicles were treated with Sema6C-Si RNA and Negative-Si RNA respectively for 72 hours, then, Lucifer Yellow microinjection and electron microscope assay were used to examine the cell junctions between granulosa cells and oocytes. We used microscope to examine their morphology including follicle atresia and development. Real-time PCR were used to determine the m RNA expression of cell junctions molecular such Connexin37??-catenin in all the groups. The related protein such Sema6C?PI3K/AKT?Connexin37??-catenin?Bax?Bcl-2 expressions were measured by western blot analysis. TUNEL assay was used to detect apoptosis of follicles. Small-interferon RNA was used to down-regulate the gene expression of Sema6 C.Results: Compared with the control group, down-regulated expression for sema6 c can defect follicular growth and increase follicle atresia, the morphology of preantral follicles has significantly changed.: the microvilli lodged and synapse between cells decreased. At the same time, m RNA and protein expression of cell juntions molecules such as Connexin37??-catenin were both decreased We also found that the expression of p-AKT has increased. Inhibited PI3K/AKT pathway can partly reverse the defect effects of low sema6 c.Conclusion: Low expression of Semaphorin6 C decreases connections between granulosa cells and oocytes to accelerate follicle atresia through PI3K/AKT pathway... |
PDF Full Text Request