Zhang Yu, Wang Qiao-Chu, Han Jun, Cao Rui, Cui Xiang-Shun, Kim Nam-Hyung, Rui Rong, Sun Shao-Chen
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, People's Republic of, China; College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, People's Republic of, China.
Mol Reprod Dev. 2014 Aug;81(8):725-34. doi: 10.1002/mrd.22341. Epub 2014 Jul 22.
Mammalian oocyte meiotic maturation involves a unique asymmetric division, but the regulatory mechanisms and signaling pathways involved are poorly understood. Dynamins are ubiquitous eukaryotic GTPases involved in membrane trafficking and actin dynamics, whose roles in mammalian oocyte maturation have not been determined. In this study, we used porcine oocytes to show that Dynamin 2 accumulated at the meiotic spindle and in the cortex of oocytes, with a distribution similar to that of actin. Inhibiting Dynamin 2 activity in porcine oocytes with the specific inhibitor dynasore resulted in failed polar-body extrusion. This phenotype may have been due to aberrant actin distribution and/or spindle positioning as inhibitor treatment disrupted the formation of the actin cap and cortical granule-free domain, which negatively impacted spindle positioning. Moreover, the distribution of ARP2, a key actin-nucleation factor, was severely reduced in the cortex after dynasore treatment. Thus, our results suggest that Dynamin 2 possibly regulates porcine oocyte maturation through its effects on actin-mediated spindle positioning and cytokinesis, and that this may depend on regulating ARP2 localization.
哺乳动物卵母细胞减数分裂成熟涉及独特的不对称分裂,但其中涉及的调控机制和信号通路尚不清楚。发动蛋白是普遍存在于真核生物中的GTP酶,参与膜运输和肌动蛋白动力学,其在哺乳动物卵母细胞成熟中的作用尚未确定。在本研究中,我们利用猪卵母细胞表明发动蛋白2在减数分裂纺锤体和卵母细胞皮质中积累,其分布与肌动蛋白相似。用特异性抑制剂dynasore抑制猪卵母细胞中的发动蛋白2活性会导致极体排出失败。这种表型可能是由于肌动蛋白分布异常和/或纺锤体定位异常,因为抑制剂处理破坏了肌动蛋白帽和无皮质颗粒区域的形成,从而对纺锤体定位产生负面影响。此外,dynasore处理后,关键肌动蛋白成核因子ARP2在皮质中的分布严重减少。因此,我们的结果表明,发动蛋白2可能通过影响肌动蛋白介导的纺锤体定位和胞质分裂来调节猪卵母细胞成熟,并且这可能依赖于调节ARP2的定位。