iTRAQ技术鉴定小鼠睾丸发育差异表达蛋白的研究

Identification for proteome profile of differentially expressed proteins in testis of comparative mouse by iTRAQ labeling

目的用同量异位素标记的相对和绝对定量(isobaric tags for relative and absolute quantitation,iTRAQ)技术结合液相色谱质谱技术构建3周龄小鼠与成年小鼠睾丸蛋白差异表达谱系。方法分别抽提3周龄小鼠与成年小鼠睾丸总蛋白质,iTRAQ标记,液相色谱(LC)分离蛋白质,LTQ OrbitrapTM质谱仪进行蛋白质鉴定。结果鉴定出20个差异表达蛋白质,其中18个为3周龄小鼠睾丸高表达蛋白质,2个为成年睾丸高表达蛋白质。差异表达蛋白质主要参与细胞的有丝分裂、减数分裂、细胞增殖、分化、凋亡及精子发生等重要细胞事件。结论用目前最新的定量蛋白质组技术,构建的3周龄小鼠睾丸与成年睾丸差异表达蛋白谱系对研究睾丸功能、精子发生有一定理论价值,为定量蛋白组学技术在雄性生殖领域的应用奠定了基础。

Objective To construct the differential expression profile of testis proteins from comparative mouse by using quantitative proteomic analysis.Methods The total protein of 3-week-old or adult mouse was extracted and labeled by isobaric tags for relative and absolute quantitation（iTRAQ）.The lebeled proteins were isolated by liquid chromatography tandem mass spectrometry（LC-MS） on linear ion trap mass spectrometer in integrated mode comprising pulsed-Q-dissociation（PQD）（LTQ OrbitrapTM） to identify the differentially expressed proteins.Results Twenty proteins were identified,among which 2 were highly expressed in the testis of adult mouse and 18 were highly expressed in 3-week-old mouse.Further bioinformatic analysis indicated that the differentially expressed proteins were related to many cell-specific processes during spermatogenesis.Conclusions By using the latest quantitative proteomic technology differentially expressed proteome profiles of testis proteins from 3 weeks and adult mouse were constructed.The profiles will inspire the further study on the function of testis and the mechanisms of spermatogenesis.