Evaluation of human sperm acrosome reastion by the activation of acroshin, and its clinical application

1. 用细胞亲和层析法纯化了具有进行性运动的人精子顶体。采用带内顶柱的连续步密度梯度法分离非顶体精子，为避免顶体反应的诱导，Percoll用不含Ca， NaHCO_3的Hanks溶液等渗。用FITC-Con A进行体化学染色，可以区分顶体反应和未反应的精子，顶体反应后暴露的顶体内膜对该剂有亲和力。在Percoll中平衡沉降表明顶体反应精子的表观密度比未反应精子轻。人类精子至少含有两种精氨酸酰胺酶。然而，上述结果表明，通过激活顶蛋白来监测顶体反应是困难的，顶蛋白与顶体内膜结合。

1. The acrosome reacted human sperm with progressive motility was purified by using cell affinity chromatography on Concanavalin A Sepharose.2. The un-acrosome reasted sperm was separated by means of the continuous-step density gradient with an inner capiral, to avoid induction of the acrosome reaction, Percoll was made isotonic with Ca, NaHCO_3 free Hanks solution.3. Hystochemical staining with FITC-Con A achieved to distinguish the acrosome reacted and un reacted sperm, the inner acrosome membrane exposed in consequence of the acrosome reaction had affinity against the agent.4. Equilibrium sedimentation in Percoll revealed that the acrosome reacted sperm had lighter apparent density than those un-reacted.5. Human sperm contained at least two arginine amidases. The above results, however, suggested it is difficult to monitor the acrosome reaction by activation of acrosin, which bound to the inner acrosome membrane.