摘要
蛋白的酪氨酸磷酸化对精子运动力的维持,精子获能,超激活运动以及顶体反应等生理过程十分重要。为探讨塔里木马鹿不同生理状态下,不同个体精液精子蛋白酪氨酸磷酸化的关系,以塔里木马鹿冷冻保存的精子为试验材料,采用非离子型去污剂(NP-40)裂解法提取精子膜蛋白,用SDS-PAGE凝胶电泳和Western免疫印迹技术检测精子膜蛋白及发生酪氨酸磷酸化蛋白的表达情况。结果表明:精液异常(黄棕色,黏稠)个体的冻融精子活力,精子蛋白酪氨酸磷酸化水平明显下降(P<0.01);含卵黄稀释液作为冷冻保存稀释时,其冻融精子蛋白的磷酸化水平显著高于无卵黄稀释液的其它个体(P<0.05);另外,相同生理状态下冷冻保存的不同个体的冻融精子酪氨酸磷酸化蛋白的表达水平也有显著差异(P<0.05),但除了精液异常个体外所有公鹿冻融精子的40 ku和47 ku蛋白的磷酸化表现出较强的表达(P<0.05)。以上结果提示,塔里木马鹿冻融精子蛋白酪氨酸磷酸化水平与精液的生理状态有关,并且不同个体之间也存在一定的差异。
Protein tyrosine phosphorylation in the maintenance of sperm motility, sperm capacitation, su- per active exercise and other physiological acrosome reaction process is very important. To explore the re- lationship of speml protein tyrosine phosphorylation with physiological states, membrane protein tyrosine phosphorylation and protein expression of thawed Tarim Wapiti sperm were detected useing non-ionic de- tergent agent （NP-40） lysis method, SDS-PAGE gel electrophoresis and Western blot. The results showed that sperm motility and protein tyrosine phosphorylation significantly decreased （ P 〈 0.01 ） in abnormal sperm （yellow brown, sticky）; diluent, protein phosphorylation levels of thawed sperm containing afrozen egg yolk dilution were significantly higher than those without yolk diluent （ P 〈 0.05） ; in addi- tion, the same physiological state cryopreserved thawed sperm of different individuals tyrosine phosphory- lated protein levels were also significantly different （ P 〈 0.05）, but in addition to abnormal sperm out- side individual all gonna freeze-thaw sperm 40 ku and 47 ku protein phosphorylation showed strong ex- pression （ P 〈 0.05）. These results suggested that there were some differences in thawed sperm protein tyrosine phosphorylation levels and the physiological state of the semen, and also between different indi- viduals in Tarim Wapiti.
出处
《经济动物学报》
CAS
2013年第3期136-139,145,共5页
Journal of Economic Animal
基金
新疆维吾尔自治区高校科研计划重点项目(XJEDU2010I24)
新疆农业大学紧缺人才专业大学生创新项目(jqzyp42011090)
关键词
塔里木马鹿
精子
蛋白酪氨酸磷酸化
差异表达
Cervus elaphus elaphus yarkandensis
sperm
protein tyrosine phosphorylation
protein ex- pression
