超低温冷冻对德国牧羊犬精子超微结构及顶体酶类活性的影响

The effect of cryopreservation on ultra-structure and acrosome enzyme activity of German shepherd dog sperm

为探讨超低温冷冻对德国牧羊犬精子超微结构及顶体酶类活性的影响,对冷冻前后德国牧羊犬精子采用扫描电镜和透射电镜观察超微结构变化;采用改良明胶底膜法、改良BNPNA法、磷酸苯二钠法对精子透明质酸酶、顶体酶和酸性磷酸酶活性进行研究。结果表明:鲜精和冻前精子形态变化不显著,而超低温冷冻后精子头部、颈部和尾部发生形态变化的精子数极显著高于鲜精和冻前精子;超低温冷冻后德国牧羊犬精子3种顶体酶活性极显著降低;超低温冷冻后德国牧羊犬精子3种顶体酶活性与精子活率和顶体完整率呈正相关(P<0.01),与畸形率呈负相关(P<0.05)。

Scanning electron microscopy （SEM） and transmission electron microscopy （TEM） were utilized to study the effect of cryopreservation on ultra-structure of shepherd dog sperm, and the enzyme activities of HYD, ACE and ACP were also determined by the modified gelatin membrane method, modified BMPNA and di-sodium phenyl phosphate. The results indicated that there was no significant change between fresh sperm and un-cryopreserved sperm in ultra-structure. However, the number of sperms with morphologically changed head, neck and/or tail in cryopreserved semen was highly significant larger （P〈0.01） than that of fresh and un-cryopreserved semen. The activities of all three acrosome enzymes were also significantly decreased （P〈0.01） due to cryopreservation. After cryopreservation, the activity of the three acrosome enzymes was positively correlated with （P〉0.01） sperm viability and acrosomal integrity respectively but negatively correlated with （P〈0.05） aberration rate.