Abstract：Proteomic changes in the semen of dairy sheep (Ovis aries) during cryopreservation may lead to sperm death or fertility decline. In order to explore the differential proteins between fresh and frozen ram semen, the semen of 18 Donghu F1 dairy sheep was collected by artificial vagina method, and frozen and preserved with glycerol and egg yolk as the main cryoprotectant. The total protein of fresh and frozen ram semen were extracted, purified, compared and analyzed by tandem mass tag (TMT) technology. The results showed that 936 proteins were screened out before and after freezing. Taking fold change (FC)>1.1 and P<0.05 as screening conditions, there were 22 differential proteins between fresh and frozen thawed semen, of which 17 proteins were up-regulated and 5 proteins were down regulated, and all of them were involved in the regulation of sperm metabolism and biological processes. GO enrichment analysis showed that 8 differential proteins were involved in the biological process and cell components. KEGG was used to analyze the signal pathways in which the differentially expressed proteins were significantly enriched. It was found that the differentially expressed proteins were mainly enriched in metabolism, protein metabolism, apoptosis and other related pathways. Two up-regulated and 2 down-regulated proteins were selected to detect their abundances in fresh and frozen semen by Western blot. The results showed that the relative abundance of the 4 proteins were similar to the results of TMT proteomics analysis, indicating that the proteomics data were reliable. This study provides basic data for screening of protein markers of semen freezing injury in Donghu F1 dairy sheep.
李德娴, 王广, 陈璐, 余梦琦, 邹家浩, 张永涛, 袁宇欣, 张磊, 李广. 东湖F1代奶绵羊精液冷冻前后的蛋白质组差异研究[J]. 农业生物技术学报, 2023, 31(3): 558-568.
LI De-Xian, WANG Guang, CHEN Lu, YU Meng-Qi, ZOU Jia-Hao, ZHANG Yong-Tao, YUAN Yu-Xin, ZHANG Lei, LI Guang. Study on Proteomic Differences of Semen Before and After Freezing in Donghu F1 Dairy Sheep (Ovis aries). 农业生物技术学报, 2023, 31(3): 558-568.
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