猪冻精繁殖生产瓶颈以及鲜精有效利用期较短已经成为限制生猪改良种育的主要原因，如何延长猪精子体外保存时间、提高优质种公猪精液利用率是目前畜牧业亟待解决的问题。前期研究结果提示猪休眠精子活力等繁殖潜能的降低与蛋白巴豆酰化修饰水平之间存在相关性。因此，本项目在猪精子休眠诱导及巴豆酰化蛋白质组学研究基础上，利用细胞亚分离技术、免疫沉淀技术、修饰肽段抗体亲和富集技术及蛋白酰化酶调控技术，分离并鉴定猪精子巴豆酰化蛋白及巴豆酰化修饰位点，定量比较新鲜精子与休眠精子蛋白巴豆酰化修饰差异性，重点发掘与受精相关巴豆酰化蛋白标记物，分析低温休眠精子蛋白巴豆酰化特点与受精功能相互关系，从精子巴豆酰化蛋白质组学角度探讨猪休眠精子受精潜能变化的分子机制，对于突破猪精子体外保存繁殖学瓶颈以及制定猪精子体外低温休眠诱导新策略具有重要科学意义和应用价值。

The bottlenecks of boar frozen semen production and the short utilization period of fresh sperm have been the main obstacles to restrict the improvement of swine breeding. Hence, extending the preservation time of boar sperm available to artificial insemination in vitro and improving the utilization rate of boar semen are urgent problems to be solved in animal husbandry. However, our previous studies suggested that the decrease of reproductive potential including motility was closely related to the protein lysine crotonylation (Kcr) levels in dormant sperm. Thus, based on the existing research of dormancy induction of boar sperm and Kcr proteomics, we use cell isolation technology, immunoprecipitation and HPLC-MS/MS to identify crotonoylated proteins as well as their Kcr sites in boar spermatozoa and meanwhile, to quantitatively analyze the difference of protein Kcr between fresh semen and dormant spermatozoa. Importantly, this project would also focus on searching the special Kcr protein markers associated with fertilization, analyzing the characteristics of protein Kcr as well as revealing the explicit relationship between protein Kcr and fertility in microthermal dormant sperm. Given all these potential findings, this project would clarify the molecular mechanisms underlying the changes of reproductive potential of dormant boar sperm from the perspective of Kcr proteomics, which will be of great scientific significance and application value to surmount the bottlenecks of boar sperm preservation and to develop new strategies for boar sperm dormancy induction at low temperature in vitro.