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

doi:10.11843/j.issn.0366-6964.2013.08.008

摘要/Abstract

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38~170 ku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。

Abstract:

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.

中图分类号:

S828
S814.8

孙培亮，崔明勋，姜园园，曹立朋，金一. 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[J]. 畜牧兽医学报, 2013, 44(8): 1229-1235.

SUN Pei-liang, CUI Ming-xun, JIANG Yuan-yuan, CAO Li-peng, JIN Yi. The Expression and Tyrosine Phosphorylation of Sperm Protein 32 Regulate the Activation of the Boar Proacrosin/Acrosin System[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(8): 1229-1235.

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