Abstract：
To investigate the mechanisms underlying the differences in the freezability of boar semen, Yorkshire boars with freezing-tolerant semen (YT, n = 3), Yorkshire boars with freezing-sensitive semen (YS, n = 3), Landrace boars with freezing-tolerant semen (LT, n = 3), and Landrace boars with freezing-sensitive semen (LS, n = 3) were selected for this study. Their sperm was subjected to protein extraction, followed by data-independent acquisition proteomics and functional bioinformatics analysis. A total of 3042 proteins were identified, of which 2810 were quantified. Some key KEGG pathways were enriched, such as starch and sucrose metabolism, carbohydrate digestion and absorption, mineral absorption, the HIF-1 signaling pathway, and the necroptosis pathways. Through PRM verification, we found that several proteins, such as α-amylase and epididymal sperm-binding protein 1, can be used as molecular markers of the freezing resistance of boar semen. Furthermore, we found that the addition of α-amylase to cryoprotective extender could significantly improve the post-thaw motility and quality of boar semen. In summary, this study revealed some molecular markers and potential molecular pathways contributing to the high or low freezability of boar sperm, identifying α-amylase as a key protein. This study is valuable for optimizing boar semen cryopreservation technology.
摘要：
为了研究猪精液冷冻能力差异的潜在机制，具有耐冻精液的约克郡公猪（YT，n=3),具有冷冻敏感性精液的约克郡公猪（YS，n=3),具有耐冻精液的长白猪（LT，n=3),和具有冷冻敏感性精液的长白猪(LS，n=3)被选择用于本研究。他们的精子经过蛋白质提取，其次是独立于数据的采集蛋白质组学和功能生物信息学分析。共鉴定出3042种蛋白质,其中有2810个被量化。丰富了一些关键的KEGG途径，如淀粉和蔗糖代谢，碳水化合物的消化和吸收，矿物质吸收，HIF-1信号通路,和坏死途径。通过PRM验证,我们发现了几种蛋白质，如α-淀粉酶和附睾精子结合蛋白1,可作为猪精液抗冻性的分子标记。此外，我们发现，在冷冻保护补充剂中添加α-淀粉酶可以显着提高猪精液的解冻后运动能力和质量。总之,这项研究揭示了一些分子标记和潜在的分子途径有助于公猪精子的高或低冷冻能力，确定α-淀粉酶为关键蛋白。本研究对优化公猪精液冷冻保存技术具有一定的参考价值。
