钙离子对猪精子蛋白酪氨酸磷酸化及活力影响  

Impact of Calcium on Protein Tyrosine Phosphorylation and Motility of Boar Sperm

作　　者：赵娜[1] 甄林青 姜红菊 严国祥 李新红[1] 

机构地区：[1]上海交通大学农业与生物学院上海市兽医生物技术重点实验室,上海200240 [2]上海市祥欣畜禽有限公司,上海201302

出　　处：《上海交通大学学报（农业科学版）》2015年第2期66-72,共7页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：农业部公益性行业专项(200903056);上海市科技兴农攻关项目(沪农科攻字2009第5-1号)

摘　　要：研究主要探究钙离子(Ca2+)对猪精子蛋白酪氨酸磷酸化及活力的作用。利用计算机辅助精子活力分析仪(CASA)检测、蛋白免疫印迹测定不同Ca2+浓度对精子活力指标及蛋白酪氨酸磷酸化水平的影响,并对精子样品的酪氨酸磷酸化蛋白的分布情况进行观察。结果显示,当Ca2+浓度≤1.0mmol/L时,精子活力和蛋白酪氨酸磷酸化水平增高;然而,当Ca2+浓度≥3.0mmol/L时,则明显抑制精子活力和蛋白酪氨酸磷酸化水平,尤其分子量约为27和34kDa的高丰度蛋白磷酸化程度明显降低(P<0.05)。高浓度Ca2+(4.0mmol/L)明显抑制鞭毛处蛋白酪氨酸磷酸化。因此,细胞外钙离子对猪精子活力和蛋白磷酸化起到重要调节作用,低浓度Ca2+促进精子活力及蛋白酪氨酸磷酸化,高浓度Ca2+(≥3.0mmol/L)抑制精子活力及蛋白酪氨酸磷酸化。本研究初步明确了Ca2+浓度对猪精子蛋白磷酸化及精子活力的影响,为进一步研究精子获能提供参考。The aim of the study was to understand the influence of calcium（Ca^2＋）on sperm protein tyrosine phosphorylation and motility in boars.Sperm motility parameters were measured using computer-assisted sperm analysis（CASA）and the level of tyrosine phosphorylation was detected by western blot.Meanwhile,distribution of tyrosine phosphorylated protein targets in boar sperm was observed using immunofluorescence technique.The results showed that the sperm motility and protein tyrosine phosphorylation were promoted by low concentrations of Ca^2＋（≤ 1.0 mmol/L）.On the contrary,at the relatively higher concentrations of Ca^2＋（≥3.0 mmol/L）,the sperm motility（MOT）and curvilinear velocity（VCL）were significantly decreased（P〈0.05）and tyrosine phosphorylation was inhibited,especially the protein phosphorylation of the molecular weight about 27 and 34kDa（P〈0.05）.The results also indicated that the distribution of tyrosine phosphorylated protein substrates was significantly reducedwith 4.0mmol/L Ca^2＋treatment.Thus,the results revealed that the changes in extracellular Ca^2＋ play a critical role in sperm motility and protein tyrosine phosphorylation.Low concentrations of calcium（≤ 1.0mmol/L）play a accelerative role on sperm motility and tyrosine phosphorylation,but the higher concentrations of calcium（≥3.0mmol/L）have a inhibitory effect.This study explicated the regulation of Ca^2＋ concentrations on boar sperm protein phosphorylation and sperm motility preliminarily,providing a useful reference for further uncovering of molecular mechanism of capacitation.

关 键 词：CA^2+ 猪 精子活力 酪氨酸磷酸化 

分 类 号：S828[农业科学—畜牧学]