C型利钠肽及其受体在水牛卵泡中的表达分析

Expression Analysis of C-type Natriuretic Peptide and Its Receptor in Buffalo Follicles

为了探明C型利钠肽(C-type natriuretic peptide,CNP)及其受体(natriuretic peptide receptor,NPR)在水牛卵泡中的表达模式,本研究首先采用实时荧光定量PCR技术检测水牛卵巢中利钠肽家族主要成员A型、B型、C型利钠肽(ANP、BNP、CNP)及其Ⅰ型、Ⅱ型受体(NPR1、NPR2)的mRNA表达情况,然后利用免疫组化技术对水牛卵泡中CNP及NPR2蛋白进行定位,最后利用实时荧光定量PCR技术检测颗粒细胞和卵丘细胞中CNP及NPR2的mRNA表达规律。结果显示,水牛卵巢主要表达CNP及NPR2,且在各级卵泡中均有表达,其中,CNP主要在壁层颗粒细胞中表达,NPR2主要在卵丘细胞中表达;颗粒细胞上CNP mRNA表达水平显著高于卵母细胞周围的卵丘细胞(P<0.05),而卵丘细胞上NPR2mRNA表达水平显著高于颗粒细胞(P<0.05)。综上所述,在利钠肽主要家族成员和受体中,CNP和NPR2在水牛卵巢中呈现强表达,CNP主要在壁层颗粒细胞中表达,而NPR2主要在卵母细胞周围的卵丘细胞中表达。

In order to investigate the expression pattern of C-type natriuretic peptide（CNP）and its receptors（NPR）in buffalo follicles,firstly,the mRNA expression level of ANP,BNP,CNP,NPR1 and NPR2in ovary were assayed by Real-time quantitative PCR;Secondly,the protein expression of CNP and NPR2 in buffalo follicles were detected by immunohistochemical stainning method;Lastly,the mRNA expression level of CNP and NPR2 in granule cells and cumulus cells were assayed by Real-time quantitative PCR.The results showed that CNPgene and its receptor NPR2 mainly expressed in buffalo ovary,the protein of CNP and NPR2 expressed in all stages of follicles,CNP mainly expressed in mural granulosa cells and NPR2 primarily presented in cumulus cells,the mRNA expression of CNP gene in granule cells was significantly higher than cumulus cells（P〈0.05）,whereas the mRNA expression of NPR2 gene in cumulus cells was significantly higher than granule cells（P〈0.05）.In conclusion,among the main members of natriuretic peptides and its receptors,CNP and NPR2 presented strong expression in buffalo ovary.CNP mainly expressed in mural granulosa cells,but NPR2 primarily presented in cumulus cells which arounding oocytes.