Shi Zhonghua, Zhao Chun, Yang Ye, Teng Hui, Guo Ying, Ma Minyue, Guo Xuejiang, Zhou Zuomin, Huo Ran, Zhou Qi
State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing, 210029, People's Republic of China.
Cell Mol Life Sci. 2015 Sep;72(18):3575-86. doi: 10.1007/s00018-015-1905-3. Epub 2015 Apr 17.
In mammals, pronucleus formation, a landmark event for egg activation and fertilization, is critical for embryonic development. However, the mechanisms underlying pronucleus formation remain unclear. Increasing evidence has shown that the transition from a mature egg to a developing embryo and the early steps of development are driven by the control of maternal cytoplasmic factors. Herein, a two-dimensional-electrophoresis-based proteomic approach was used in metaphase II and parthenogenetically activated mouse eggs to search for maternal proteins involved in egg activation, one of which was poly(rC)-binding protein 1 (PCBP1). Phosphoprotein staining indicated that PCBP1 displayed dephosphorylation in parthenogenetically activated egg, which possibly boosts its ability to bind to mRNAs. We identified 75 mRNAs expressed in mouse eggs that contained the characteristic PCBP1-binding CU-rich sequence in the 3'-UTR. Among them, we focused on H2a.x mRNA, as it was closely related to pronucleus formation in Xenopus oocytes. Further studies suggested that PCBP1 could bind to H2a.x mRNA and enhance its stability, thus promoting mouse pronucleus formation during parthenogenetic activation of murine eggs, while the inhibition of PCBP1 evidently retarded pronucleus formation. In summary, these data propose that PCBP1 may serve as a novel maternal factor that is required for determining the normal timing of pronucleus formation.
在哺乳动物中,原核形成是卵子激活和受精的标志性事件,对胚胎发育至关重要。然而,原核形成的潜在机制仍不清楚。越来越多的证据表明,从成熟卵子到发育中胚胎的转变以及发育的早期步骤是由母体细胞质因子的控制驱动的。在此,基于二维电泳的蛋白质组学方法被用于中期II和孤雌激活的小鼠卵子,以寻找参与卵子激活的母体蛋白,其中之一是多聚(rC)结合蛋白1(PCBP1)。磷酸蛋白染色表明,PCBP1在孤雌激活的卵子中表现出去磷酸化,这可能增强其与mRNA结合的能力。我们鉴定出在小鼠卵子中表达的75种mRNA,它们在3'-UTR中含有特征性的PCBP1结合富含CU的序列。其中,我们重点关注H2a.x mRNA,因为它与非洲爪蟾卵母细胞中的原核形成密切相关。进一步的研究表明,PCBP1可以结合H2a.x mRNA并增强其稳定性,从而在小鼠卵子的孤雌激活过程中促进小鼠原核形成,而抑制PCBP1明显延迟原核形成。总之,这些数据表明PCBP1可能作为一种新的母体因子,是决定原核形成正常时间所必需的。