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目的:鉴定大鼠附睾管腔液中可能参与精子成熟/精子表面修饰的附睾特异蛋白。方法:通过SDS-PAGE或双向(2D)电泳比较正常大鼠附睾头段和尾段以及实验性精索静脉曲张(ELV)大鼠附睾尾段管腔液中差异蛋白成分。选择主要差异斑点进行质谱分析,并采用免疫印迹对目标蛋白作进一步鉴定确认。结果:发现多个蛋白成分明显增高。对其中之一的相对分子质量(Mr)22000蛋白集中研究。经质谱分析及免疫印迹确认Mr22000蛋白为磷脂酰乙醇胺结合蛋白(PBP)。结论:附睾管腔液中的PBP存在多种分子形式包括糖基化修饰。ELV导致大鼠附睾尾管腔液中PBP升高。因此,该分子与精子表面修饰的关系仍需进一步研究。
OBJECTIVE: To identify epididymal specific proteins that may participate in sperm maturation / sperm surface modification in rat epididymal fluid. Methods: The differential proteins in the caudal and caudal segments of the epididymis and the cauda equina of experimental varicocele (ELV) rats were compared by SDS-PAGE or two-dimensional electrophoresis. The major difference spots were selected for mass spectrometry analysis and further identification of the target protein was performed using immunoblotting. Results: Multiple protein components were found to be significantly higher. One of these studies focused on the relative molecular mass (Mr) 22000 protein. The Mr22000 protein was identified as phosphatidylethanolamine binding protein (PBP) by mass spectrometry and Western blotting. Conclusion: There are many molecular forms of PBP in epididymal fluid including glycosylation. ELV causes elevated PBP in the rat epididymal lumen. Therefore, the relationship between the molecule and sperm surface modification needs further study.