Zhang Xiao-Lan, Liu Peng, Yang Zhi-Xia, Zhao Jing-Jing, Gao Lei-Lei, Yuan Bo, Shi Li-Ya, Zhou Chun-Xiang, Qiao Hai-Feng, Liu Ya-Hong, Ying Xiao-Yan, Zhang Jun-Qiang, Ling Xiu-Feng, Zhang Dong
State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China.
Department of Veterinary Drug Supervision, Taiyuan, Shanxi, China.
Oncotarget. 2017 Jun 9;8(57):96809-96825. doi: 10.18632/oncotarget.18425. eCollection 2017 Nov 14.
PNMA (paraneoplastic antigen MA) family includes Pnma1-6. Although other members have been found to be involved in paraneoplastic neurological disorders, death receptor-dependent apoptosis, and tumorigenesis, Pnma5 was thought to be a female fertility factor, as indicated by one genome-wide study. But until now there have not been any further functional studies about Pnma5 in female meiosis. Our preliminary study indicated that Pnma5 might play important roles in meiosis. To further address this, Pnma5 was knocked down in in-vitro maturated (IVM) mouse oocytes, which are common models for mammalian female meiosis, by specific siRNA, and results showed that the loss of Pnma5 significantly delayed the progression of meiosis I and increased chromosome segregation errors during anaphase I. In in-vitro fertilization (IVF), Pnma5 knockdown caused significantly lower fertilization. To assess how it affects meiosis, Pnma5 knockdown was found to significantly decrease the stability of spindle microtubules and altered F-actin organization within actin cap regions, cause significantly abnormal mitochondria aggregation and lower ATP concentration. Next we have found that phosphorylation at Thr533 re-located Pnma5 strongly to spindles & cortex and was required for the phosphorylation of Akt and Gsk3β, while Src and Erk1/2 phosphorylation was required for the phosphorylation of Pnma5, indicating that phosphorylated Pnma5 is the active form and subsequently activates Akt and Gsk3β. Collectively this study suggests that Pnma5 is important for meiosis and is the pivot of Src→Erk1/2→Pnma5→Akt→Gsk3β pathway.
副肿瘤抗原MA(PNMA)家族包括Pnma1 - 6。尽管已发现其他成员与副肿瘤性神经系统疾病、死亡受体依赖性细胞凋亡和肿瘤发生有关,但一项全基因组研究表明,Pnma5被认为是一种女性生育因子。但迄今为止,尚未有关于Pnma5在雌性减数分裂中功能的进一步研究。我们的初步研究表明,Pnma5可能在减数分裂中发挥重要作用。为进一步探究此问题,通过特异性小干扰RNA(siRNA)在体外成熟(IVM)的小鼠卵母细胞(哺乳动物雌性减数分裂的常用模型)中敲低Pnma5,结果显示,Pnma5的缺失显著延迟了减数分裂I的进程,并增加了后期I染色体分离错误。在体外受精(IVF)中,Pnma5敲低导致受精率显著降低。为评估其对减数分裂的影响,发现Pnma5敲低显著降低了纺锤体微管的稳定性,并改变了肌动蛋白帽区域内的F - 肌动蛋白组织,导致线粒体聚集明显异常且ATP浓度降低。接下来我们发现,苏氨酸533位点的磷酸化使Pnma5强烈重新定位于纺锤体和皮质,这是Akt和Gsk3β磷酸化所必需的,而Src和Erk1/2磷酸化是Pnma5磷酸化所必需的,表明磷酸化的Pnma5是活性形式,随后激活Akt和Gsk3β。总体而言,这项研究表明Pnma5对减数分裂很重要,并且是Src→Erk1/2→Pnma5→Akt→Gsk3β信号通路的枢纽。