斑马鱼早期胚胎发育囊胚sphere时期的蛋白组学研究

斑马鱼(Danio rerio)早期胚胎发育囊胚时期是胚胎发育过程的关键时期,利用i TRAQ蛋白质质谱分析技术,检测斑马鱼早期胚胎发育过程中囊胚sphere时期的蛋白质表达情况,并分析该时期表达的蛋白质的相应功能和参与调控的生物过程。以野生型斑马鱼发育至sphere时期即4 hpf的胚胎为样本,利用i TRAQ标记与LC-MS/MS串联质谱技术,结合数据库比对,对该时期表达的蛋白质进行定性和定量的鉴定分析。检测结果共鉴定到的总蛋白数为1 178个,利用生物信息学进行功能分析,发现这些蛋白广泛参与了细胞信号传递、细胞运动和细胞骨架构建、细胞增殖、细胞分化、物质合成与代谢等各项重要的生命活动过程。研究表明,利用i TRAQ标记的方法可以对4hpf时期的斑马鱼胚胎中的蛋白质进行有效的分离和鉴定,并初步建立了斑马鱼早期发育关键阶段sphere时期的蛋白质组表达图谱,以期为斑马鱼胚胎发育过程中蛋白质组学和调控机制的研究提供参考。

Analyses of proteomic profile at the embryonic developmental sphere stage of Danio rerio embryos

The blastula stage of zebrafish in early embryogenesis is an important process.iTRAQ protein mass spectrometry was used to detect the protein expression profile at blastula transition sphere stage in zebrafish early embryogenesis,and to reveal the function of the proteins and the biological process involved at this stage.The embryos of wild-type zebrafish at sphere stage(4hpf)were used as protein sample,and by iTRAQ labeling combined with LC-MS/MS analysis method and comparison with zebrafish protein database the proteins were qualitatively and quantitatively identified at this stage.A total of 1 178 proteins were identified,and further bioinformatics analysis revealed that these proteins were mostly involved in many biological processes which were essential for development,such as cell proliferation,cell differentiation,material synthesis and metabolism,cellular signal and transferring,cell migration and cytoskeleton,etc.In this study,the proteins expression at 4hpf stage of zebrafish embryos were successfully separated and identified by iTRAQ method.Preliminarily the proteomic profile was established at the critical sphere embryonic developmental stage of zebrafish,and it will provide references for further proteomic and regulatory mechanism research in zebrafish embryogenesis.