水牛成熟前后卵泡液的多肽组鉴定和分析

Identification and Analysis of Peptidome from Buffalo Mature and Immature Follicular Fluid

卵泡液为雌性配子的成熟和发育提供生长微环境,在不同卵泡发育时期,卵泡液中的蛋白质成分会产生变化,这些变化与卵泡发育、排卵和卵母细胞成熟有直接关系。本研究对水牛卵泡液开展多肽组学分析,采用超滤方法富集内源性多肽,建立了稳定的多肽组制备方法。使用MALDI-TOF飞行时间质谱仪采集多肽信号,共采集到297个质谱峰,鉴定得到43个多肽或蛋白质前体,95%的多肽分子量在15 kD以下。其中成熟卵泡液鉴定得到36种,未成熟卵泡液中鉴定得到24种,两者均能鉴定得到17种。经生物信息学分析,在鉴定得到的多肽中,共参与细胞因子受体相互作用、细胞周期和生长激素信号传导等7个调控代谢通路。本实验的多肽组学研究为卵泡发育机理提供了新的研究方法和参考数据。

Follicular fluid provided the microenviroment for oocyte maturation and development. The components of proteins in follicular fluid are variable in different development period. All the proteins changes were related to folliculo genesis, ovulation and oocyte maturation. In this study, follicular fluid peptidomics analysis was performed on buffalo. Endogenous peptidome were enriched by ultrafiltration method and peptidome samples preparation method was established. Matrix-assisted Laser Desorption Ionization Time of Flight（MALDI-TOF） mass spectrometer was utilized to acquire the peptides signal, a total of 297 mass spectrums were obtained from mature and immature follicular fluid, all the mass spectrums were corresponding to 43 peptides or protein precursors. 95% of peptides molecular weight（MW） were below 15 kD. 36 peptides were identified from mature follicular fluid, 24 peptides were identified from immature follicular fluid. While a total of 17 peptides were identified in both groups. The result of bioinformatical analysis indicated that all the peptides and protein precursors were involved in seven regulation pathways including Cytokine-Cytokine receptor interaction, cell cycle and GnRH signaling pathway. The result of peptidome analysis provided a new approach and a dataset for the study of folliculogenesis mechanism.