Wang Ningling, Zhang Ping, Guo Xuejiang, Xie Jin, Huo Ran, Wang Fuqiang, Chen Lin, Shen Jian, Zhou Zuomin, Shi Qinghua, Zhao Baige, Sha Jiahao
Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing, P. R. China.
Proteomics. 2009 Jul;9(13):3425-34. doi: 10.1002/pmic.200800822.
The assembly of primordial follicles early in ovarian development and subsequent transition to primary follicles are critical processes in ovarian biology. Inappropriate coordination of these processes contributes to ovarian pathologies such as premature ovarian failure and infertility. To better understand the molecular mechanisms involved in primordial follicle assembly and development, 2-D PAGE and MALDI-TOF/TOF technologies were used to construct a comparative proteome profile of the immature rat ovary at specific time-points (0, 24, 48, and 72 h postpartum). A total of 154 differential protein spots corresponding to 134 different proteins were definitively identified between any two time-points. Further cluster analysis showed four expression patterns, and each pattern correlated with specific cell processes that occur during early ovarian development. Seven proteins were randomly selected to verify expression patterns using Western blotting, and subsequently immunohistochemistry was performed to further investigate their cellular localization. Additionally, detailed functional analyses of these differentially expressed proteins were performed. Elucidation of how these changes in protein expression level coordinate primordial follicles assembly and development is intended to provide a better understanding of these critical biological processes early in ovarian development and will provide potential therapeutic molecular targets to regulate ovarian function and treat ovarian disease.
卵巢发育早期原始卵泡的组装以及随后向初级卵泡的转变是卵巢生物学中的关键过程。这些过程的协调不当会导致卵巢疾病,如卵巢早衰和不孕。为了更好地理解原始卵泡组装和发育所涉及的分子机制,采用二维聚丙烯酰胺凝胶电泳(2-D PAGE)和基质辅助激光解吸电离飞行时间串联质谱(MALDI-TOF/TOF)技术构建了产后特定时间点(0、24、48和72小时)未成熟大鼠卵巢的比较蛋白质组图谱。在任意两个时间点之间共明确鉴定出154个差异蛋白点,对应134种不同蛋白质。进一步的聚类分析显示出四种表达模式,每种模式都与卵巢早期发育过程中发生的特定细胞过程相关。随机选择七种蛋白质,使用蛋白质印迹法验证表达模式,随后进行免疫组织化学以进一步研究它们的细胞定位。此外,对这些差异表达蛋白质进行了详细的功能分析。阐明蛋白质表达水平的这些变化如何协调原始卵泡的组装和发育,旨在更好地理解卵巢发育早期的这些关键生物学过程,并将为调节卵巢功能和治疗卵巢疾病提供潜在的治疗分子靶点。