间隙连接蛋白Cx43和Cx45在体外受精绵羊早期胚胎中的表达

为了研究间隙连接蛋白Cx43和Cx45在体外受精绵羊胚胎早期发育过程各阶段的表达情况,利用RT-PCR扩增得到绵羊Cx43(43 ku)及Cx45(45 ku)基因的mRNA,并进行序列测定。收集绵羊未成熟和成熟卵母细胞以及体外受精早期胚胎各发育阶段的卵裂球细胞,通过RT-PCR半定量检测Cx43和Cx45基因的mRNA表达量;通过免疫组织化学方法结合激光扫描共聚焦显微镜检测Cx43和Cx45蛋白在绵羊体外受精的早期胚胎发育过程中的表达情况及表达区域。结果表明,在绵羊卵母细胞成熟过程以及早期胚胎发育的各个时期,Cx43及Cx45在mRNA水平和蛋白水平均有表达;Cx43、Cx45蛋白主要分布在细胞膜表面,在细胞质和细胞核中表达微弱。研究结果为进一步探索Cx43和Cx45在绵羊早期胚胎发育过程中的作用提供了试验数据。

Expression of connexin 43 and connexin 45 in sheep preimplantation embryos fertilized in vitro

To study the expression of connexin43(Cx43) and connexin45(Cx45) in sheep preimplantation embryos,RT-PCR was used to detect the mRNA level of the 2 connexins, and their cDNA were sequenced.Ovine immatured oocytes,matured oocytes and embryos from 2-cell stage to blastocyst produced in vitro were recovered,and RT-PCR was used to semi-quantify mRNA expression.Proteins of Cx43 and Cx45 were detected by using a combination of immunocytochemistry and Laser Scanning Confocal Microscope.Cx43 and Cx45 were expressed in oocytes and preimplantation embryos at mRNA and protein le-(vels.) The 2 connexins distributed mainly on the surface of cell membrane and could not be detected in the nuclei and cytoplasm.The results provided foundation for future studies of functions of Cx43 and Cx45 in sheep preimplantation embryos.