杜洛克公猪精液NAGase对调控精子功能蛋白质组学的研究  

Research on proteomics of NAGase in regulating the function of spermatozoa in Duroc boar semen

作　　者：林秋敏[1,2] 赖宝色[1] 钟佩云 林祚贵 徐磊[1] 黄小红[2] LIN Qiumin;LAI Baose;ZHONG Peiyun;LIN Zuogui;XU Lei;HUANG Xiaohong(College of Animal Science and Technology,Fujian Vocational College of Agricultural,Fuzhou 350007,China;Fujian Province Key Laboratory of Integrated Traditional Chinese and Western Veterinary Medicine and Animal Health Care,Fujian Agriculture and Forestry University,Fuzhou 350002,China)

机构地区：[1]福建农业职业技术学院动物科技学院,福建福州350007 [2]福建农林大学中西兽医结合与动物保健福建省高校重点实验室,福建福州350002

出　　处：《畜牧与兽医》2025年第2期14-21,共8页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金项目(31572484);福建农业职业技术学院博士科研启动课题(2022JS002);2023年福建农业职业技术学院科技研究项目(2023JS002)。

摘　　要：旨在以杜洛克公猪精液为研究对象,通过N-乙酰-β-D-氨基葡萄糖苷酶(NAGase)抑制剂体外处理精液,经12.5%SDS-PAGE对处理好的精子样品进行双向电泳分离,筛选NAGase作用互作点蛋白表达差异;表达丰度差异的蛋白经酶消化后进行质谱分析。结果:银染筛选获得57个差异表达蛋白点在凝胶中,蛋白点主要分布在pH值3~10和相对分子质量14.4~116 kDa的范围内;经NCBI和Mascot数据库进行差异表达蛋白匹配搜索,成功鉴定与精子功能相关的上调差异表达蛋白8个,分别是磷脂酰乙醇胺结合蛋白4(PEBP4)、精卵融合蛋白4(IZUMO4)、精浆糖蛋白1(PSP-1)、猪精子黏附蛋白(AWN)、糖结合蛋白(SPMI)、细胞色素c氧化酶Ⅵα亚基多肽1(COX6A1)、二氢硫辛酸脱氢酶蛋白(DLD)和伴侣蛋白1(TCP1);8个上调差异表达蛋白主要涉及精子成熟、精子运动、精卵识别等生理功能。通过STRING工具分析蛋白质互作关系表明IZUMO蛋白可能是NAGase调节精子功能的一个重要靶点并形成了可能的蛋白质调控网络。研究结果为提高公猪生产性能研究奠定了基础。The aim of this study was to investigate the semen of Duroc boars,specifically by treating the semen with an N-acetyl-β-Dglucosaminidase(NAGase)inhibitor in vitro.Following this treatment,the processed sperm samples were subjected to two-dimensional electrophoresis separation using 12.5%SDS-PAGE to screen for differences in NAGase-interacting proteins.Finally,proteins exhibiting abundance differences were digested with enzymes and were analyzed by mass spectrometry.The results were as follows:A total of 57 differentially expressed protein spots were detected in the gel after silver staining,with the protein spots primarily distributed within a pH range of 3 to 10 and a relative molecular mass range of 14.4 to 116 kDa.Differential protein matching searches conducted using NCBI and Mascot databases successfully identified eight upregulated differential proteins related to sperm function,namely phosphatidylethanolamine-binding protein 4(PEBP4),IZUMO sperm-egg fusion protein 4(IZUMO4),glycoprotein in seminal plasma 1(PSP-1),porcine sperm adhesion protein(AWN),sugar-binding protein(SPMI),cytochrome C oxidase VI A subunit polypeptide 1(COX6A1),dihydrolipoyl dehydrogenase protein(DLD),and chaperonin protein 1(TCP1).These eight upregulated differentially expressed proteins were primarily involved in physiological functions such as sperm maturation,sperm motility,and sperm-egg recognition.Analysis of protein-protein interactions using the STRING tool indicated that the IZUMO protein might be an important target for NAGase in regulating sperm function,forming a potential protein regulatory network.The findings of this study provided a basis for animal reproduction and production and laid the groundwork for further research on improving animal production performance.

关 键 词：杜洛克公猪 N-乙酰-Β-D-氨基葡萄糖苷酶 精子功能 蛋白质组学 

分 类 号：S828[农业科学—畜牧学]