| Connexin43 is the most expressed connexin in granulosa cells of mammalian ovarian follicles and plays an important role in follicular development and oocyte growth.The expression of Cx43 in granulosa cells is regulated by gonadotropins,it has been found that follicle stimulating hormone(FSH)can promote the expression of Cx43 in granulosa cells of developing follicles and strengthen the gap junction between granulosa cells.Whether luteinizing hormone(LH)can promote the expression of Cx43 and its specific mechanism is not clear.In addition,androgen plays an important role in the process of follicular development,and the effect of androgen on the expression of Cx43 in granulosa cells is still unknow.This study use the ovary from different developmental stages of female C57 mice and the primary cultured rat granulosa cells for materials,application of morphology,cell biology and molecular biology methods,to study the expression pattern of Cx43 during the growth of mice,effects of luteinizing hormone(LH)and dihydrotestosterone(DHT)on the expression of Cx43 in rat granulosa cells and the possible mechanism.The results of this study showed that Cx43 was expressed in cell membrane of granulosa and LHR was expressed in the cytoplasm of the theca cells,granulosa cells and oocytes.The expression of Cx43 and LHR increased significantly with the follicular development and the expression of Cx43 was significantly correlated with LHR.Intervention of LH and DHT increased the expression of Cx43 in ovarian tissues.After incubated with different concentrations of LH and DHT in vitro,we found that intervention of these two moleculesincreased the expression of Cx43 in mouse ovary.To 21 days old rat granulosa cells,the results showed that Cx43 expressed in cytoplasm and the part of membrane in granulosa cells.LH and LiCl(agonist of Wnt/β-catenin signaling pathway)promoted the expression of Cx43,While FH535(inhibitor of Wnt/β-catenin signaling pathway)could decrease the Cx43 expression induced by LH;similarly,DHT and LiCl(agonist of Wnt/β-catenin signaling pathway)promoted the expression of Cx43,While FH535(inhibitor of Wnt/β-catenin signaling pathway)could decrease the Cx43 expression induced by DHT.In conclusion:(1)In the state of mice natural growth,the expression of Cx43 and LHR were significantly increased with follicular development.LH could regulate the expression of Cx43 in granulosa cells through LHR.(2)Suitable concentrations of LH and DHT incubated the mouse ovarian tissue can promote the expression of Cx43 in granulosa cells,indicating that LH and DHT are the regulatory factors of Cx43 expression.(3)LH and DHT stimulated the expression of Cx43 in granulosa cells by activing the Wnt/β-catenin signaling pathway. |
