【作者】 库尔班·吐拉克； 陈勇； 王旭光； 迪力夏提； 敬斌宇； 李和平；

【机构】 东北林业大学野生动物资源学院； 新疆农业大学动物科学学院； 新疆维吾尔自治区畜牧总站； 新疆厚拾生物科技有限责任公司；

【摘要】 为探讨不同获能方法对塔里木马鹿精子体外获能及其蛋白酪氨酸磷酸化水平的影响,冻融塔里木马鹿精子随机分为4组,用钙离子载体、肝素、咖啡因和Percool离心4种方法进行精子获能的诱导,利用金霉素(CTC)染色法评价精子获能状态,采用SDS-PAGE分离精子膜蛋白,进行Western blot免疫印迹分析,检测酪氨酸磷酸化蛋白的表达水平。结果显示,冻融精子经4种精子获能方法处理后,肝素诱发的精子获能率显著高于IA组和Percool组(P<0.05)。IA组、肝素组和咖啡因组精子蛋白酪氨酸磷酸化水平高于Percool组和对照组。另外,冻融精子随着上游处理及肝素诱导获能的进行,检测到分子量分别为14ku、25~30ku、40 ku、47 ku、55 ku的酪氨酸磷酸化蛋白,这些蛋白的酪氨酸磷酸化水平在获能60~120min期间相对较高,而且此时精子获能率及超激活运动精子比例也显著提高(P<0.05)。结果提示,肝素可以较好地诱导马鹿精子获能,马鹿精子获能与蛋白酪氨酸磷酸化相关。更多还原

【Abstract】 In this paper,the effects of different treatment on eapaeitation and protein tyrosine phosphorylation of Tarim Wapiti sperm were studied.Frozen- thawed Tarim Wapiti sperms were divided into 4 groups randomly and treated with A23187(IA),Heparin,caffeine or Pereool for eapaeitation.The sperm eapaeitation was evaluated by Aureomyein(CTC)method.Expression of sperm protein tyrosine phosphorylation was detected by membrane- protein separation on SDS- PAGE and Western blotting subsequently.The results indicated that sperm eapaeitation rate in treatment of heparin- induced was significantly higher than the IA treatment and Percool treatment(P < 0.05).The expression of sperm protein tyrosine phosphorylation in each group of IA,heparin and caffeine was increased than the Percool treatment and the control.With swim- up and heparin- induced,the expression of 14 ku,25- 30 ku,40 ku,47 ku and 55 ku protein tyrosine phosphorylation was detected subsequently,these proteins tyrosine phosphorylation were increased when the frozen- thawed sperms capacitated 60 ~120mins,and the capacitated sperm rate and hyper-activated sperm rate were increased significantly(P < 0.05).The results suggested that sperms of Tarim Wapiti were capacitated properly with heparin and protein tyrosine phosphorylation was associated with the eapaeitation of Tarim Wapiti sperm.更多还原