Cheng Jia, Hao Xu, Zhang Weijing, Sun Chenhao, Yuan Xiameng, Yang Yiding, Zeng Wenxian, Zhu Zhendong
School of Biological Science and Engineering, Shaanxi University of Technology, Hanzhong 723001, China.
Qinba Mountain Area Collaborative Innovation Center of Bio-Resources Comprehensive Development, Hanzhong 723001, China.
Animals (Basel). 2024 Dec 19;14(24):3672. doi: 10.3390/ani14243672.
Sex-controlled sperm combined with artificial insemination allows animals to reproduce offspring according to the desired sex, accelerates the process of animal genetics and breeding and promotes the development of animal husbandry. However, the molecular markers for sexual sperm sorting are unusual. To identify the molecular markers of boar sperm sorting, proteomics and metabolomics techniques were applied to analyze the differences in proteins and metabolism between X and Y sperm. Label-free quantitative proteomics identified 254 differentially expressed proteins (DEPs) in the X and Y sperm of boars, including 106 proteins that were highly expressed in X sperm and 148 proteins that were highly expressed in Y sperm. Among the differential proteins, COX6A1, COX1, CYTB, FUT8, GSTK1 and PFK1 were selected as potential biological markers for X and Y sperm sorting. Moreover, 760 metabolites from X and Y sperm were detected. There were 439 positive ion mode metabolites and 321 negative ion mode metabolites identified. The various metabolites were phosphoenolpyruvate, phytosphingosine, L-arginine, N-acetylputrescine, cytidine-5'-diphosphate and deoxyuridine. These metabolites were mainly involved in the TCA cycle, oxidative phosphorylation pathway, glycolysis pathway, lipid metabolism pathway, amino acid metabolism pathway, pentose phosphate pathway and nucleic acid metabolism pathway. The differential proteins and differential metabolites obtained by the combined proteomics and metabolomics analysis were projected simultaneously to the KEGG pathway, and a total of five pathways were enriched, namely oxidative phosphorylation pathway, purine metabolism, unsaturated fatty acid biosynthesis, ABC transporters and peroxisomes. In summary, COX6A1 and CYTB were identified as potential biomarkers for boar X and Y sperm sorting.
性控精子结合人工授精可使动物按期望的性别繁殖后代，加速动物遗传育种进程，推动畜牧业发展。然而，用于精子性别分选的分子标记并不常见。为鉴定公猪精子分选的分子标记，应用蛋白质组学和代谢组学技术分析X和Y精子之间蛋白质和代谢的差异。无标记定量蛋白质组学鉴定出公猪X和Y精子中有254种差异表达蛋白（DEPs），其中106种蛋白在X精子中高表达，148种蛋白在Y精子中高表达。在差异蛋白中，COX6A1、COX1、CYTB、FUT8、GSTK1和PFK1被选为X和Y精子分选的潜在生物学标记。此外，检测到X和Y精子中的760种代谢物。鉴定出439种正离子模式代谢物和321种负离子模式代谢物。这些代谢物包括磷酸烯醇丙酮酸、植物鞘氨醇、L-精氨酸、N-乙酰腐胺、胞苷-5'-二磷酸和脱氧尿苷。这些代谢物主要参与三羧酸循环、氧化磷酸化途径、糖酵解途径、脂质代谢途径、氨基酸代谢途径、磷酸戊糖途径和核酸代谢途径。通过蛋白质组学和代谢组学联合分析获得的差异蛋白和差异代谢物同时投射到KEGG通路，共富集了五条通路，即氧化磷酸化途径、嘌呤代谢、不饱和脂肪酸生物合成、ABC转运蛋白和过氧化物酶体。综上所述，COX6A1和CYTB被鉴定为公猪X和Y精子分选的潜在生物标志物。
