College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China.
Biol Reprod. 2019 Mar 1;100(3):711-720. doi: 10.1093/biolre/ioy217.
Actin filaments are widely involved in multiple cellular processes in oocyte meiosis, such as spindle migration and polar body extrusion. The actin nucleators like Arp2/3 complex and formins are the most recognized molecules for actin assembly in oocytes. In the present study, we report that the vesicle trafficking factor, RAB8A GTPase, is a new regulator critical for actin assembly in meiosis. Our results showed that RAB8A was localized at both the spindle periphery and cortex in mouse oocytes, which was similar to the localization patterns of actin filaments. RAB8A depletion caused spindle migration defects and the failure of polar body extrusion, which could have been due to decreases in both cytoplasmic and cortical actin filaments in oocytes. Based on mass spectrometry analysis, we showed that RAB8A promoted actin assembly through its modulation on the ROCK-LIMK signaling pathway. Moreover, we demonstrated that RAB8A colocalized and interacted with GM130 at the spindle periphery and that RAB8A depletion caused the disruption of GM130-docked Golgi distribution. Taken together, our data indicated that RAB8A was required for Golgi distribution, spindle migration, and polar body extrusion via ROCK-mediated actin assembly in mouse oocyte meiosis.
肌动蛋白丝广泛参与卵母细胞减数分裂中的多种细胞过程,如纺锤体迁移和极体排出。肌动蛋白成核因子如 Arp2/3 复合物和formin 是卵母细胞中肌动蛋白组装最被认可的分子。在本研究中,我们报告了囊泡运输因子 RAB8A GTPase 是新的关键调节因子,对于减数分裂中的肌动蛋白组装至关重要。我们的结果表明,RAB8A 定位于小鼠卵母细胞的纺锤体周围和皮质,与肌动蛋白丝的定位模式相似。RAB8A 的耗竭导致纺锤体迁移缺陷和极体排出失败,这可能是由于卵母细胞中细胞质和皮质肌动蛋白丝的减少。基于质谱分析,我们表明 RAB8A 通过调节 ROCK-LIMK 信号通路促进肌动蛋白组装。此外,我们证明 RAB8A 在纺锤体周围与 GM130 共定位并相互作用,并且 RAB8A 的耗竭导致 GM130 停靠的高尔基体分布中断。总之,我们的数据表明,RAB8A 通过 ROCK 介导的肌动蛋白组装在小鼠卵母细胞减数分裂中对于高尔基体分布、纺锤体迁移和极体排出是必需的。