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

作　　者：毛飞 杨燕燕[2] 苏杰 赵高平 李秀男[1,2] 王大清 巴音吉日嘎拉 曹贵方[1,3] 王彩云[1] MAO Fei;YANG Yanyan;SU Jie;ZHAO Gaoping;LI Xiunan;WANG Daqing;Bayinjrigala;CAO Guifang;WANG Caiyun(College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China;Animal Husbandry Institute,Inner Mongolia Academy of Agricultural&Animal Husbandry Sciences,Hohhot 010031,China;Inner Mongolia Saikexing Institute of Domestic Animal Breeding and Reproduction Biotechnology,Hohhot 011517,China)

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018 [2]内蒙古农牧业科学院畜牧研究所,内蒙古呼和浩特010031 [3]内蒙古赛科星家畜种业与繁育生物技术研究院,内蒙古呼和浩特011517

出　　处：《畜牧与兽医》2023年第9期10-16,共7页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金项目(31860689);内蒙古自治区科技计划项目(2019GG241);数字成像技术用于多脊椎项目(2020GG0055);内蒙古自治区科技成果转化引导项目(2020CG0078);肉羊冷冻精液制作关键技术应用研究项目(2019GG348);奶牛良种关键技术开发项目(2021GG0403);内蒙古农业大学教改项目(RC2100002194)。

摘　　要：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.

关 键 词：蛋白质组学 精子获能 乳铁蛋白 绵羊 

分 类 号：S826[农业科学—畜牧学]