牛精子蛋白质组学技术平台的建立及冻融前后精子差异蛋白初步分析

本研究通过探索不同的精子蛋白制备方法、水化液成分和优化2D电泳程序以建立牛精子蛋白质组学研究技术平台，同时以牛鲜冻精为实验材料通过差异凝胶电泳寻找冻融前后精子蛋白的改变。结果表明：使用改进的热TRIzol法裂解精子细胞制备蛋白，结合优化的2D电泳技术可建立稳定的牛精子蛋白质组学研究技术平台。差异凝胶电泳揭示牛精子在冻融后有质和量的改变：冻融后缺失的蛋白点有20个，表达下调的有2个，表达上调的有10个。作为一项阶段性的实验成果，本研究建立的2D平台和所发现的冻融引起的差异表达蛋白质点为揭示冷冻损伤机理和性控精液的差异蛋白质组学研究奠定了较好的基础。

Establishment of proteomics methods of bovine spermatozoa and initial analysis of the differential proteins between the fresh and frozen-thawed spermatozoa

Abstract The proteomics technique platform of bovine spermatozoa was established by exploring the different sample preparation methods, the components of rehydration solution and optimizing the 2D electrophoresis protocols. The differential proteins between the fresh and frozen-thawed spermatozoa were also explored. The results indicated that the technique system with the high qualitative and stable 2D image had been obtained by preparing the bovine spermatozoa proteins with the heat TRIzol method and combining with the optimized 2D electrophoresis protocols. The differential in-gel electrophoresis showed that 20 protein spots were absent in quality, 2 spots decreased and 10 spots increased significantly in quantity after frozen-thawed. As a intermediate result, the 2D platform established in the research and the differential proteins found between fresh and frozen sperm will provide better basis for revealing the cryodamage mechanism of sperm cryopreservation and studying the differential proteomics of the X, Y spermatozoa.