精子蛋白32的表达及酪氨酸磷酸化调控猪精子顶体蛋白的活化  

The Expression and Tyrosine Phosphorylation of Sperm Protein 32 Regulate the Activation of the Boar Proacrosin/Acrosin System

作　　者：孙培亮[1] 崔明勋[1] 姜园园[1] 曹立朋[1] 金一[1] 

机构地区：[1]延边大学农学院,延吉133002

出　　处：《畜牧兽医学报》2013年第8期1229-1235,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31260529)

摘　　要：为了探究猪精子蛋白32(Sperm protein,sp32)表达及酪氨酸磷酸化调控猪精子顶体蛋白活化的关系,本研究对不同处理(鲜精、冷冻-解冻、获能、顶体反应)的猪精子顶体膜蛋白进行分离,通过考马斯亮蓝染色、SDS-PAGE电泳和Western blot检测和分析。结果表明,猪精子经过获能处理、冷冻-解冻处理以及顶体反应处理后,前顶体蛋白(Proacrosin)和顶体蛋白(Acrosin)在转化过程中,sp32表达有所差异,获能和顶体反应处理的sp32的表达量略高于冷冻-解冻处理组,而显著高于鲜精处理组。与其他处理组相比,冷冻-解冻精子处理组sp32酪氨酸磷酸化水平产生显著的差异。但在SDS-PAGE电泳中,猪鲜精处理组在蛋白质分子质量38～170ku区间的蛋白表达条带较其他对比组明显,这表明猪精子在受精前所必需经历的获能和顶体反应过程中,顶体膜蛋白伴随着大分子的蛋白质修饰和降解。结论:sp32作为一种前顶体蛋白结合蛋白,在前顶体蛋白活化过程中它的表达水平及酪氨酸磷酸化水平上调。The aim of this study was to investigate the relationship between the expression level,tyrosine phosphorylation of sp32 and the activation of the boar proacrosin/acrosin system,the acrosomal membrane proteins of boar sperm for different treatments（fresh sperm,freezing-thawing,capacitation,acrosome reaction） were separated,and stained by CBB,assayed using SDS-PAGE and Western blot analysis.The results showed that the expression level of sp32 was different in conversion of proacrosin/acrosin system after treatment of boar sperm capacitation,freezing-thawing and acrosome reaction,the expression of sp32 in experiment groups of capacitation and acrosome reaction were slightly higher than the freezing-thawing experiment group,and significantly higher than that of fresh semen group.The level of sp32 tyrosine phosphorylation had a significant difference between the freezing and thawing experiment group and other experiment groups.But bands with molecular mass of 38-170 ku in the fresh semen group were more obvious;it showed that the acrosomal membrane proteins were accompanied by modification and degradation of big molecular proteins when sperm underwent capacitation and the acrosome reaction.sp32 as a proacrosin binding protein,its expression and tyrosine phosphorylation level were up-regulated in the activation of the boar proacrosin/acrosin system.

关 键 词：sp32 前顶体蛋白 顶体蛋白 活化 酪氨酸磷酸化 

分 类 号：S828[农业科学—畜牧学] S814.8[农业科学—畜牧兽医]