猪精子获能期间蛋白质磷酸化时间依赖性的上调  

Time-dependent Upreulation of Protein Phosphorylation during Sperm Capacitation in Pigs

作　　者：张旭成[1] 袁慧敏[1] 金一[1] 

机构地区：[1]延边大学农学院,吉林龙井133400

出　　处：《江西农业大学学报》2010年第3期577-580,共4页Acta Agriculturae Universitatis Jiangxiensis

基　　金：国家自然科学基金项目(30960250)

摘　　要：通过获能猪精子酪氨酸磷酸化蛋白分离和表达,来确定精子获能的最佳状态。将猪精子悬浮于改良的Tris缓冲液(mTBM)获能培养基中,在体积分数为5%CO2孵箱37℃培养,以考马斯亮蓝染液染色的方法来评价精子获能状态,经过SDS-PAGE分离后,进行Western免疫印迹分析,检测获能猪精子间酪氨酸磷酸化蛋白的分布。有27,47 ku的2种蛋白发生酪氨酸磷酸化,其中27 ku的蛋白酪氨酸磷酸化水平自精子体外培养后呈递增趋势,而47 ku的蛋白酪氨酸磷酸化水平在精子培养1.5 h时最高,后呈递减趋势,1.5 h时获能状态最佳。To determine the best condition for sperm capacitating by the separation and expression of tyrosin phoshporylated protein.Sperm from mature pigs was incubated in modified mTBM under 5% CO2 in air at 37 ℃.The effect of capacitation was assessed by Coomassie brilliant blue staining.Using Western blotting analysis after SDS-PAGE separation the distribution of tyrosine phosphorylated protein of pig sperm of capacitation was determined.There were two proteins（27 ku,47 ku）phosphorylated in this experiment.The protein of 27 ku was detected by anti-phosphotyrosine monoclonal antibody,and the intensity of this protein increased in incubation.Then another protein of 47 ku was found and the level of this protein reached the highest point at 1.5 h,and the intensity of the protein decreasing in incubation.1.5 h is the best condition for sperm capacitation.

关 键 词：猪 酪氨酸磷酸化 精子获能 SDS-PAGE电泳 WESTERN印迹 

分 类 号：S828.11[农业科学—畜牧学]