促性腺激素调控半滑舌鳎(Cynoglossus semilaevis)卵母细胞孕酮受体膜组分1的表达特征

摘要

In this study we investigated the characteristics and mechanisms of the expression of progesterone receptor membrane component 1 (PGRMC1) during oocyte maturation of the half-smooth tongue sole (Cynoglossus semilaevis).Expression of PGRMC1 mRNA in the oogenesis of the ovarian development stages Ⅳ and Ⅴ were monitored with qRT-PCR technique.The results showed that the highest level of PGRMC1 mRNA appeared in Phase Ⅴ oocytes of the mature C.semilaevis.In general,the transcription of PGRMC1 peaked in different developmental phases of oocyte.This suggested that PGRMC1 participated in ovarian development and maturation of C.semilaevis.Next we tested whether HCG treatment affected PGRMC 1 expression in different stages of oogenesis using qRT-PCR and western blotting analysis.It was shown that HCG could up-regulate the expression of PGRMC1 mRNA and protein,and that 20 IU/ml HCG was more powerful than 10 IU/ml (P＜0.05).The effect of HCG varied in different phases of oocyte and was the most obvious in Phase Ⅴ,indicating that PGRMC1 played an important role in the mature stage.The results above suggested that PGRMC1 could regulate the maturation of C.semilaevis oocytes.Our findings may shed light on future exploration of the role of PGRMC 1 in the reproductive function of C.semilaevis.%采用qRT-PCR方法分析性成熟半滑舌鳎(Cynoglossus semilaevis)不同发育时期卵巢卵母细胞的孕酮受体膜组分1 (PGRMC1)mRNA的表达,研究发现,在发育Ⅳ期的卵巢,处于第Ⅳ时相的卵母细胞PGRMC1 mRNA表达量最高(P＜0.05);在发育Ⅴ期的卵巢,成熟期卵母细胞的PGRMC1mRNA表达量最高(P＜0.05).采用不同浓度促性腺激素(HCG)处理卵巢发育Ⅴ期半滑舌鳎不同时相的卵母细胞,并通过qRT-PCR和Western blotting技术对其表达量变化进行检测.结果显示,20IU/mlHCG对PGRMC1 mRNA和蛋白表达的调控作用比10 IU/ml HCG作用更明显,表明PGRMC1 mRNA和蛋白表达对HCG调控作用存在剂量依存关系.HCG对半滑舌鳎不同时相的卵母细胞的调控作用效果不同,对第Ⅴ时相卵母细胞作用最明显(P＜0.05),表明PGRMC1主要在卵母细胞成熟阶段发挥作用.PGRMC1对HCG调控作用的正向应答效应预示其参与了卵母细胞成熟调控,为进一步探讨PGRMC1在半滑舌鳎繁殖过程的功能提供重要基础资料.