获能前后绵羊精子的差异蛋白质组学分析

Differential proteome analysis of the sperm of sheep before and after capacitation

为了探索获能前后绵羊精子蛋白质的动态变化,通过串联质谱标签结合液质联用分析技术,对获能前后绵羊精子的总蛋白进行定量分析。结果显示:获能前后绵羊精子的差异表达蛋白共有203个,获能后表达上调蛋白质27个,表达下调蛋白质176个。差异表达参与解剖结构发育、生殖等生物学进程;从细胞组分看,差异表达蛋白定位在细胞溶质、质膜中;从分子功能看,主要参与氧化还原等过程。差异表达蛋白也参与蛋白酶体、Hedgehog和cAMP等信号通路。通过差异蛋白表达结合文献分析,筛选出乳铁蛋白。Western blot结果表明,乳铁蛋白在绵羊精子中存在,且在获能后表达量极显著降低(P<0.0001);RT-qPCR结果表明,乳铁蛋白在获能前后绵羊精子中均存在,且差异极显著(P<0.001),说明乳铁蛋白对绵羊精子获能有一定的影响。

In order to explore the dynamic changes in protein in sheep sperm before and after capacitation,the total protein in the sheep sperm before and after capacitation was quantitatively analyzed using tandem mass spectrometry label combined with liquid chromatographymass spectrometry.The results showed that there were 203 dfferentially expressed proteins in the sperm before and after capacitation,with 27 up-regulated proteins and 176 down-regulated proteins after capacitation.Diferential expression was involved in the biological processes,such as anatomical structure development and reproduction.In terms of cell components,the differentially expressed proteins were located in the solutes and plasma membranes of the cells.In the perspective of molecular functions,the proteins were mainly involved in the redox processes.The differentially expressed proteins were also involved in the proteasome,Hedgehog and cAMP signaling pathways.Lactoferrin was screened by differential protein table combined with literature analysis.The results of Western blot showed that lactoferrin existed in the sheep sperm and the expression level decreased after capacitation(P<0.0001).The results of RT-qPCR showed that lactoferrin existed in both the sperm of the control group and the capacitation group(P<0.001).In short,lactoferrin had a certain effect on sheep sperm capacitation.