Centre de recherche en biologie de la reproduction, Département des sciences animales, FSAA, Université Laval, Québec, Québec, Canada.
PLoS One. 2013 Jul 4;8(7):e68456. doi: 10.1371/journal.pone.0068456. Print 2013.
Gap junctional communication (GJC) plays a primordial role in oocyte maturation and meiotic resumption in mammals by directing the transfer of numerous molecules between cumulus cells and the oocyte. Gap junctions are made of connexins (Cx), proteins that regulate GJC in numerous ways. Understanding the dynamic regulation of connexin arrangements during in vitro maturation (IVM) could provide a powerful tool for controlling meiotic resumption and consequently in vitro development of fully competent oocytes. However, physiological events happening during the early hours of IVM may still be elucidated. The present study reports the dynamic regulation of connexin expression, degradation and localization during this stage. Cx43, Cx45 and Cx60 were identified as the main connexins expressed in swine COC. Cx43 and Cx45 transcripts were judged too static to be a regulator of GJC, while Cx43 protein expression was highly responsive to gonadotropins, suggesting that it might be the principal regulator of GJC. In addition, the degradation of Cx43 expressed after 4.5 h of IVM in response to equine chorionic gonadotropin appeared to involve the proteasomal complex. Cx43 localisation appeared to be associated with GJC. Taken together, these results show for the first time that gonadotropins regulate Cx43 protein expression, degradation and localisation in porcine COC during the first several hours of IVM. Regulation of Cx43 may in turn, via GJC, participate in the development of fully competent oocytes.
缝隙连接通讯(GJC)通过指导卵丘细胞和卵母细胞之间大量分子的转移,在哺乳动物卵母细胞成熟和减数分裂恢复中发挥着首要作用。缝隙连接由连接蛋白(Cx)组成,这些蛋白通过多种方式调节 GJC。了解连接蛋白排列在体外成熟(IVM)过程中的动态调节,可能为控制减数分裂恢复提供有力工具,从而控制完全有能力的卵母细胞的体外发育。然而,IVM 早期发生的生理事件仍有待阐明。本研究报告了在这一阶段连接蛋白表达、降解和定位的动态调节。Cx43、Cx45 和 Cx60 被鉴定为猪 COC 中主要表达的连接蛋白。Cx43 和 Cx45 转录本被认为过于静态,不能作为 GJC 的调节剂,而 Cx43 蛋白表达对促性腺激素高度敏感,表明它可能是 GJC 的主要调节剂。此外,在 IVM 4.5 小时后,Cx43 的降解似乎涉及蛋白酶体复合物。Cx43 的定位似乎与 GJC 有关。综上所述,这些结果首次表明,在 IVM 的最初几个小时内,促性腺激素调节猪 COC 中 Cx43 蛋白的表达、降解和定位。Cx43 的调节可能通过 GJC 参与完全有能力的卵母细胞的发育。