mPRα在性成熟雌性牙鲆(Paralichthys olivaceus)不同组织中的定性定量表达特征

摘要

In this study we collected the mature tissue samples of female Japanese Flounder Paralichthys olivaceus and investigated the expression patterns of mRNA and proteins of membrane progestin receptor alpha (mPRα) in different tissues and different oocyte phases,using techniques such as quantitative real-time PCR,in situ hybridization,immunohistochemistry,and westem blotting.It was shown that the highest level of mPRα mRNA appeared in the Ⅴ phase of oocyte maturation.This further implied that mPRα was involved in the regulation of oocyte maturation through guiding progesterone.The western blotting analysis confirmed the expression of mPRα proteins in the ovary,brain,head kidney,kidney,and liver.Moreover,the expression of mPRα mRNA and proteins was higher in the brain and ovary than in other tissues.These results indicated that mPRα could play certain roles in different tissues,especially the endocrine-related ones.Results of in situ hybridization and immunohistochemistry clearly showed that the mPRα mRNA and proteins were expressed on the oocyte membrane and the bureaucratic structures of other organs.Our study enriched the knowledge about mechanisms of membrane receptor signal transduction.%通过荧光实时定量PCR方法(qRT-PCR)检测雌性性成熟牙鲆(Paralichthys olivaceus)第Ⅴ时期不同时相的卵母细胞中膜孕激素受体(mPRα)的表达,同时采用原位杂交、免疫组化和Western blotting方法研究性成熟雌性牙鲆mPRα mRNA和蛋白在各组织中的分布和表达.qRT-PCR结果显示,在牙鲆卵巢成熟时期的第Ⅴ时相卵母细胞,mPRα mRNA的表达量最高,表明mPRα介导孕激素参与了牙鲆卵母细胞成熟的调控.组织定量定位结果显示,在牙鲆的卵巢、脑、头肾、肾、肝脏组织中均有mPRα mRNA和蛋白表达存在,且在脑、卵巢中表达量较高,表明mPRα在牙鲆不同组织中都发挥着一定的作用,且在内分泌相关组织如脑和卵巢中的作用更明显.原位杂交和免疫组化结果显示,mPRα mRNA和蛋白表达明显定位在牙鲆卵母细胞膜上,且分布在其他组织的管腔结构.本研究为进一步探究牙鲆mPRα信号转导机制提供了理论基础和形态学依据.