不同耐冻性荣昌猪精子冻后质量及靶向脂质组学分析  

Post-Freezing Quality and Targeted Lipidomics Analysis of Rongchang Pig Spermatozoa with Different Freezing Tolerance

作　　者：高小平 潘红梅[1,4] 郭宗义 张军杰[2] 林燕 张亮 GAO XiaoPing;PAN HongMei;GUO ZongYi;ZHANG JunJie;LIN Yan;ZHANG Liang(Chongqing Academic of Animal Science,Chongqing 402460;College of Life Sciences,Sichuan Agricultural University,Ya’an 625000,Sichuan;Department of Animal Nutrition Institute,Sichuan Agricultural University,Chengdu 215500;Pig Technology Innovation Center(Chongqing),Chongqing 402460)

机构地区：[1]重庆市畜牧科学院,重庆402460 [2]四川农业大学生命科学学院,四川雅安625000 [3]四川农业大学动物营养研究所,成都611100 [4]生猪技术创新中心(重庆),重庆402460

出　　处：《中国农业科学》2025年第2期387-400,I0012-I0016,共19页Scientia Agricultura Sinica

基　　金：国家重点研发计划课题(2021YFD1200303);四川省生猪创新团队项目(SCCXTD-2024-8);国家生猪技术创新中心先导科技项目(NCTIP-XD/B04)。

摘　　要：【背景】荣昌猪是我国优良地方猪种之一,但随着外种猪的引入以及市场需求的转变,纯种荣昌猪的数量正不断下降,这一趋势严重威胁到了我国生猪遗传资源的多样性和养猪业的可持续发展。在地方猪冷冻精液制作过程中,不同公猪精子冻后质量的差异都与其耐冻性有关,而精子脂质组成是影响其耐冻性的关键因素之一。【目的】通过深入分析不同耐冻性荣昌猪精子的质量特征及其脂质组成,筛选出精子耐冻性的候选标志物。为猪精液冷冻保存研究提供依据。【方法】采集14头荣昌猪精液进行冷冻保存后检测其冻后活力,以相对活力(冻后活力/冻前活力)为耐冻性标准,对荣昌猪精子进行高、低耐冻性筛选并分组。检测冻后不同耐冻组精子的质膜完整性、线粒体膜电位、活性氧水平、凋亡水平和体外卵母细胞穿透能力,并通过扫描电镜和透射电镜观察精子表面和内部微观结构的冷冻损伤程度。通过靶向脂质组学技术检测不同耐冻组新鲜精子的中长链脂肪酸组成,筛选出差异脂肪酸。【结果】通过相对活力筛选出具有高耐冻性精子荣昌猪5头,具有低耐冻性精子荣昌猪5头;冷冻后高耐冻组精子的质膜完整性、线粒体活性和体外穿卵率显著高于低耐冻组精子(P<0.05);冷冻后高耐冻组精子凋亡率显著低于低耐冻组精子(P<0.05)。冷冻后高耐冻组精子表面和内部结构完整性均高于低耐组精子。靶向脂质组学显示,荣昌猪精子中含有36种脂肪酸。高、低耐冻组精子间棕榈酸、十五烷酸等11种脂肪酸含量存在显著差异(P<0.05),且均在高耐冻组中更丰富。【结论】荣昌猪高耐冻性精子在生理状态、形态结构以及体外卵母细胞穿透功能方面都显著优于低耐冻性精子,且脂肪酸组成存在显著差异。本研究筛选出的差异脂肪酸如棕榈酸、十五烷酸、花生四烯酸等可作为荣昌猪�【Background】Rongchang pig is one of the excellent local pig breeds in China,but the number of purebred Rongchang pigs is decreasing with the introduction of foreign-bred pigs as well as the shift of the market demand,and this trend seriously threatens the diversity of the genetic resources of pigs in China and the sustainable development of the pig breeding industry.In the process of frozen semen production in local pigs,the differences in post-freezing quality of spermatozoa from different boars are all related to their freezing tolerances,while the sperm lipid composition is one of the key factors affecting their freezing tolerance.【Objective】The aim of this study was deeply analyze the quality characteristics of spermatozoa of Rongchang pigs with different freezing tolerances and their lipid compositions,and to screen out the candidate markers of sperm freezing tolerance.【Method】Fourteen Rongchang pig semen were collected for freezing and preservation,and their post-freezing motilities were detected.Relative Motility(post-freezing motility/pre-freezing motility)was used as the criterion for freezing tolerance,and Rongchang pig spermatozoa were screened and grouped into groups for high and low freezing tolerance.Plasma membrane integrity,mitochondrial membrane potential,reactive oxygen species level,apoptosis level and in vitro oocyte penetration ability of spermatozoa from different freezing tolerance groups were detected after freezing,and the extent of freezing damages on the surface and internal microstructure of spermatozoa were observed by scanning electron microscopy and transmission electron microscopy.The medium-and long-chain fatty acid composition of fresh spermatozoa from different freeze-resistant groups was detected by targeted lipidomics technology,and differential fatty acids were screened out.【Result】5 Rongchang pigs with high freezing tolerance spermatozoa and 5 Rongchang pigs with low freezing tolerance spermatozoa were screened by relative motility;the plasma membrane integri

关 键 词：荣昌猪 精子耐冻性 靶向脂质组学 

分 类 号：S828[农业科学—畜牧学]