Localization of rabbit sperm acrosin during the acrosome reaction induced by immobilized zona matrix.
Localization of rabbit sperm acrosin during the acrosome reaction induced by immobilized zona matrix.
复制标题
固定化透明带基质诱导顶体反应期间兔精子顶体蛋白的定位。
DOI：
10.1095/biolreprod45.1.20
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发表时间：
1991
影响因子：
3.6
通讯作者：
O'Rand,MG
中科院分区：
文献类型：
--
作者：
Richardson,RT;Nikolajczyk,BS;Abdullah,LH;Beavers,JC;O'Rand,MG
为了更好地了解顶体帽的损失的表面上的透明带和赤道顶体后区域的顶体反应后的功能，我们已经构建了一个体外系统，使用热溶解透明带干燥到盖玻片上，并与获能精子孵育。该系统可以对精子与透明带的相互作用进行良好的光学分辨率。通过盖玻片上的透明带诱导的顶体反应（30%）与先前报道的使用溶解的透明带溶液诱导兔精子的反应相当。兔顶体酶原抗血清，猪49 kDa的顶体酶原片段的抗血清，和猪14 kDa的C-末端顶体酶片段的抗血清被用来监测顶体反应。兔顶体酶原/顶体酶不存在于活的、顶体完整的、游动的精子表面。在与抗体接触后，顶体反应开始，顶体蛋白原/顶体蛋白酶变得可用于结合抗体，首先在顶端区域呈新月形，然后更靠后，直到整个前顶体被标记。顶体蛋白酶原/顶体蛋白酶保留在顶体反应的精子的赤道区和顶体后区，并且即使在精子不再与顶体帽相关联之后也仍然与顶体帽相关联。
To better understand the loss of the acrosomal cap on the surface of the zona pellucida and the function of the equatorial-postacrosomal region after the acrosome reaction, we have constructed an in vitro system using heat-solubilized zonae pellucidae dried onto a coverslip and incubated with capacitated spermatozoa. This system allows good optical resolution of spermatozoonzona interaction. Induction of the acrosome reaction by zonae on coverslips (30%) is comparable to the induction of the reaction reported previously for rabbit spermatozoa using solubilized zonae in solution. Antiserum to rabbit proacrosin, antiserum to a porcine 49-kDa proacrosin fragment, and antiserum to a porcine 14-kDa C-terminal acrosin fragment were utilized to monitor the acrosome reaction. Rabbit proacrosin/acrosin is not present on the surface of live, acrosome-intact, swimming spermatozoa. After contact with zona, the acrosome reaction begins and proacrosin/acrosin becomes available to bind antibody, first as a crescent in the apical region and then more posteriorly until the entire anterior acrosome is labeled. Proacrosin/acrosin remains on the equatorial and postacrosomal regions of acrosome-reacted spermatozoa and also remains associated with the acrosomal cap even after the spermatozoon is no longer associated with it. Further studies using zona-coated coverslips should lead to a more detailed understanding of the mechanism of zona penetration.