Department of Urology, Pediatric Urology and Andrology, Justus Liebig University of Giessen, Rudolf-Buchheim-Strasse 7, Germany.
Biosci Rep. 2009 Dec 15;30(3):193-200. doi: 10.1042/BSR20090088.
The eukaryotic VDAC (voltage-dependent anion channel) is a pore-forming protein originally discovered in the outer membrane of mitochondria. It has been established as a key player in mitochondrial metabolism and ion signalling. In addition, in recent years, it has also been proposed that VDAC is present in extra-mitochondrial membranes, and it has been related to cytoskeletal structures. However, little is known about the presence and intracellular localization of VDAC subtypes in mammalian gametes. In the present study, we confirm the synthesis of VDAC1 and 2 subtypes in GV (germinal vesicle) and MII (meiosis II) stage porcine oocytes as well as their protein expression. A shift in the abundance of immunoreactive 32 kDa VDAC protein between GV and MII stage oocytes was observed with anti-VDAC2 antibody. Furthermore, subcellular localization by confocal laser microscopy demonstrated fluorescent labelling of VDAC1 over the entire oocyte surface, suggesting the presence of VDAC1 in the porcine oocyte plasma membrane and around the cortical area. Anti-VDAC2 immunostaining yielded ring-like clusters of structures distributed on the cortical area in some GV, but not in MII, stage oocytes. These results are the first data obtained for VDAC in mammalian female gametes and provide the basis for studying protein-protein interactions, distribution and possible functions of VDAC subtypes during maturation and fertilization of mammalian oocytes.
真核 VDAC(电压依赖性阴离子通道)是一种最初在线粒体的外膜中发现的孔形成蛋白。它已被确立为线粒体代谢和离子信号的关键参与者。此外,近年来,还提出 VDAC 存在于线粒体以外的膜中,并与细胞骨架结构有关。然而,对于哺乳动物配子中 VDAC 亚型的存在和细胞内定位知之甚少。在本研究中,我们证实了 VDAC1 和 2 亚型在 GV(生发泡)和 MII(第二次减数分裂)期猪卵母细胞中的合成及其蛋白表达。用抗 VDAC2 抗体观察到 GV 和 MII 期卵母细胞之间 32 kDa 免疫反应性 VDAC 蛋白丰度的变化。此外,通过共聚焦激光显微镜进行的亚细胞定位显示 VDAC1 在整个卵母细胞表面的荧光标记,表明 VDAC1 存在于猪卵母细胞的质膜和皮质区域周围。抗 VDAC2 免疫染色在一些 GV 期卵母细胞中产生分布在皮质区域的环状簇状结构,但在 MII 期卵母细胞中没有。这些结果是哺乳动物雌性配子中 VDAC 的首批数据,为研究蛋白质-蛋白质相互作用、分布以及哺乳动物卵母细胞成熟和受精过程中 VDAC 亚型的可能功能提供了基础。