Zhang Ping, Ni Xiaojian, Guo Ying, Guo Xuejiang, Wang Yufeng, Zhou Zuomin, Huo Ran, Sha Jiahao
Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing, PR China.
BMC Genomics. 2009 Aug 3;10:348. doi: 10.1186/1471-2164-10-348.
The mature mouse oocyte contains the full complement of maternal proteins required for fertilization, reprogramming, zygotic gene activation (ZGA), and the early stages of embryogenesis. However, due to limitations of traditional proteomics strategies, only a few abundantly expressed proteins have yet been identified. Our laboratory applied a more effective strategy: one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (1D SDS-PAGE) and reverse-phase liquid chromatography tandem mass spectrometry (RP-LC-MS/MS) were employed to analyze the mature oocyte proteome in depth.
Using this high-performance proteomic approach, we successfully identified 625 different proteins from 2700 mature mouse oocytes lacking zona pellucidae. This is the largest catalog of mature mouse oocyte proteins compiled to date. According to their pattern of expression, we screened 76 maternal proteins with high levels of mRNA expression both in oocytes and fertilized eggs. Many well-known maternal effect proteins were included in this subset, including MATER and NPM2. In addition, our mouse oocyte proteome was compared with a recently published mouse embryonic stem cell (ESC) proteome and 371 overlapping proteins were identified.
This proteomics analysis will be a valuable resource to aid in the characterization of important maternal proteins involved in oogenesis, fertilization, early embryonic development and in revealing their mechanisms of action.
成熟的小鼠卵母细胞含有受精、重编程、合子基因激活(ZGA)以及胚胎发生早期阶段所需的全部母体蛋白质。然而,由于传统蛋白质组学策略的局限性,目前仅鉴定出少数高表达的蛋白质。我们实验室采用了一种更有效的策略:利用一维十二烷基硫酸钠聚丙烯酰胺凝胶电泳(1D SDS-PAGE)和反相液相色谱串联质谱(RP-LC-MS/MS)对成熟卵母细胞蛋白质组进行深入分析。
通过这种高性能蛋白质组学方法,我们成功地从2700个去除透明带的成熟小鼠卵母细胞中鉴定出625种不同的蛋白质。这是迄今编制的最大的成熟小鼠卵母细胞蛋白质目录。根据它们的表达模式,我们筛选出了76种在卵母细胞和受精卵中均有高水平mRNA表达的母体蛋白质。这个子集中包含了许多著名的母体效应蛋白,包括MATER和NPM2。此外,我们将小鼠卵母细胞蛋白质组与最近发表的小鼠胚胎干细胞(ESC)蛋白质组进行了比较,鉴定出371种重叠蛋白。
这种蛋白质组学分析将成为一种有价值的资源,有助于表征参与卵子发生、受精、早期胚胎发育的重要母体蛋白质,并揭示其作用机制。