Zhang Yu, Duan Xing, Xiong Bo, Cui Xiang-Shun, Kim Nam-Hyung, Rui Rong, Sun Shao-Chen
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, China; College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China.
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, China.
Theriogenology. 2014 Jul 1;82(1):49-56. doi: 10.1016/j.theriogenology.2014.02.020. Epub 2014 Mar 2.
The inhibitor Y-27632 is a specific selective inhibitor of Rho-associated protein kinases (ROCKs), which are downstream effectors of Rho guanosine triphosphatease (GTPases) and regulate Rho-associated cellular functions, including actin cytoskeletal organization. Little is known regarding the effects of Y-27632 on mammalian oocyte maturation. In the present study, we investigated the effects of Y-27632 on porcine oocyte meiosis and possible regulatory mechanisms of ROCK during porcine oocyte maturation. We found that ROCK accumulated not only at spindles, but also at the cortex in porcine oocytes. Y-27632 treatment reduced ROCK expression, and inhibited porcine oocyte meiotic maturation, which might be because of the impairment of actin expression and actin-related spindle positioning. Y-27632 treatment also disrupted the formation of actin cap and cortical granule-free domain, which further confirmed a spindle positioning failure. Thus, Y-27632 has significant effects on the meiotic competence of mammalian oocytes by reducing ROCK expression, and the regulation is related to its effects on actin-mediated spindle positioning.
抑制剂Y-27632是Rho相关蛋白激酶(ROCKs)的特异性选择性抑制剂,Rho相关蛋白激酶是Rho鸟苷三磷酸酶(GTPases)的下游效应器,可调节Rho相关的细胞功能,包括肌动蛋白细胞骨架组织。关于Y-27632对哺乳动物卵母细胞成熟的影响知之甚少。在本研究中,我们研究了Y-27632对猪卵母细胞减数分裂的影响以及ROCK在猪卵母细胞成熟过程中的可能调控机制。我们发现ROCK不仅在纺锤体上积累,而且在猪卵母细胞的皮质中积累。Y-27632处理降低了ROCK表达,并抑制了猪卵母细胞的减数分裂成熟,这可能是由于肌动蛋白表达和肌动蛋白相关纺锤体定位受损所致。Y-27632处理还破坏了肌动蛋白帽和无皮质颗粒区域的形成,这进一步证实了纺锤体定位失败。因此,Y-27632通过降低ROCK表达对哺乳动物卵母细胞的减数分裂能力有显著影响,并且这种调节与其对肌动蛋白介导的纺锤体定位的影响有关。