中国荷斯坦公牛不同耐冻性精子的蛋白质组学分析

doi:10.11843/j.issn.0366-6964.2024.03.018

摘要/Abstract

摘要： 在以冷冻精液和人工授精为主导技术的奶牛繁育体系中，公牛精液的冻后活力备受关注。精子的耐冻性是影响精液冻后活力的关键。已有研究发现解冻后的精子活力在不同公牛之间存在显著差异。因此，迫切需要从遗传水平挖掘影响精子耐冻性的关键基因和分子标记，为提高我国种公牛的精子耐冻性提供理论依据。本研究筛选了精子高耐冻组公牛9头（鲜精活力 >0.65，冻后活力 >0.40）和精子低耐冻组公牛6头（鲜精活力 >0.65，冻后活力 ≤0.27），使用Label-free蛋白质组学技术对15头公牛的精子细胞进行定量蛋白质组学分析和生物信息学分析。结果显示，在高、低耐冻组间共检测到432个差异表达蛋白，主要富集在"代谢途径"和"氧化磷酸化"等与精子能量供应密切相关的生物学过程。差异表达蛋白互作网络分析显示与能量代谢相关的蛋白之间具有强相互作用。与QTL数据库比对，发现9个差异表达蛋白与已定位到的冻后活精子百分比性状关联基因重叠。最终筛选到HSPA1A、BSP3、ACSL4等重要候选蛋白可作为公牛精子耐冻性的潜在生物标志物。本研究揭示了具有不同耐冻性公牛精子的蛋白质组特征，这些发现为我国种公牛冻后精液品质的分子选育提供了重要信息。

关键词: 精子, 耐冻性, 中国荷斯坦公牛, 蛋白质组, 功能基因

Abstract: In dairy cattle breeding systems dominated by cryopreserved semen and artificial insemination, the post-freezing motility of bull sperm is of great interest. Sperm freezability is the key to the post-freezing sperm motility. Previous studies have shown significant differences in sperm freezability among different bulls. Therefore, it is necessary to explore genes and molecular markers that affect sperm freezing tolerance at the genetic level, in order to lay a foundation for improving the sperm freezability of Chinese bulls. The present study investigated the sperm proteome of Holstein bulls in the high sperm freezability group (n=9) and the low sperm freezability group (n=6). The fresh sperm motility of bulls in the high sperm freezability group was greater than 0.65 and post-freezing motility greater than 0.4, while the fresh sperm motility of bulls in the low sperm freezability group was greater than 0.65 and post-freezing motility less than 0.27. Quantitative proteomic analysis was conducted on sperm cells of 15 bulls using label-free proteomics technology followed by bioinformatics analysis. As a result, a total of 432 differentially expressed proteins were identified between the high and low freezability groups, which were mainly enriched in biological processes closely related to sperm energy supply, such as metabolic pathway and oxidative phosphorylation. The analysis of protein-protein interaction networks showed that there were strong interactions between proteins related to energy metabolism. Compared with the QTL database, it was found that 9 differentially expressed proteins overlapped with genes associated with percentage live sperms after thawing. Overall, several important candidate proteins were identified as potential biomarkers for the freezing resistance of bull sperms, such as HSPA1A, BSP3 and ACSL4. This study revealed the proteomic characteristics of bull sperms with different freezability, which provides important information for molecular breeding of frozen semen quality in Chinese bulls.

Key words: sperm, freezability, Chinese Holstein bulls, proteome, functional genes

中图分类号:

S823.3

曹晋康, 张纯, 王佳瑶, 李晓彤, 王鹏宇, 方颖妍, 张昱, 丁宁, 姜力. 中国荷斯坦公牛不同耐冻性精子的蛋白质组学分析[J]. 畜牧兽医学报, 2024, 55(3): 1052-1061.

CAO Jinkang, ZHANG Chun, WANG Jiayao, LI Xiaotong, WANG Pengyu, FANG Yingyan, ZHANG Yu, DING Ning, JIANG Li. Proteomic Analysis of Sperm with Different Freezability in Chinese Holstein Bulls[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1052-1061.

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