OEP、SDS与卵黄的相互作用时间及温度对犬冻融精子质量的影响

Effects of Different Treatments of OEP,SDS and Egg-Yolk on Characteristics of Canines Frozen-Thawed Spermatozoa

人工采取8只优质犬精液,以果糖-Tris-柠檬酸所组成的稀释液为基础,分别于试验前0.5 h和24 h将OEP,SDS和卵黄混合于基础稀释液中,并于5℃冰箱、22℃两个温度条件下保存,用时离心选取上清液作为稀释剂并制成细管冻精.利用流式细胞术和高倍显微镜对犬冻融精子活力、质膜完整率和顶体完整率等特性进行测定,目的在于比较不同温度条件下OEP,SDS和卵黄相互作用时间的长短对犬冻融精子质量的影响,以探讨OEP,SDS对冻融精子的保护作用机理.添加稀释液前24 h混合OEP和卵黄有利于维持冻融过程中质膜完整率(61.5%)和顶体完整率(66.8%),显著提高冻融精子的总活性比率(20.2%),对犬冻融精子的保护效果明显好于相互作用0.5 h实验组的保护效果(P<0.05);OEP,SDS与卵黄相互作用时的温度对犬冻融精子质量无显著影响(P>0.05),OEP对犬冻融精子的保护效果明显优于SDS.结果提示增加OEP,SDS与卵黄相互作用的时间可以提高犬冻融精子的质量.

The basic extender of Tris-Citric add dilution was added with different concentrations of OEP,SD8 and eggyolk respectively in 0.5 and 24 h before experiment, these two groups were preserved in 5 and 22 ℃ respectively. Semen of eight high quality dogs was diluted with supernatant fluid into 8 groups and preserved in straw. Sperm motility, plasma membrane integrity and acrosomal integrity of frozen-thawed spermatozoa were evaluated using flow eytometry and high power microscope to compare the effects of the interactional time of OEP and SI38 with egg-yolk in different temperature on the quality of dog spermatozoa after freezing-thawing. Results were as following： mixing OEP and egg-yolk in 24 h before added with extender was useful to plasma membrane integrity （61.5 % ） and acrosomal integrity （66.8 % ） of frozenthawed spermatoeoa, the total motility raised prominent （20.2%） and the effect of protection to frozen-thawed sprermatozoa was better than the experimental groups of interactional time was 0.5 h （ P〈 0.05）. The temperature of adding OEP and SI38 to egg-yolk had no prominent effect on the quality of frozen-thawed sprermatozoa （P〈0.05）. Comparing with SI38, the protecting effect of OEP on frozen-thawed sprermatozoa was excelled obviously. The result showed that increasing the treating time of OEP and SI38 interacting with egg-yolk could increase the quality of frozen-thawed sprermatozxoa.