Jaiswal B S, Eisenbach M, Tur-Kaspa I
Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot, Israel.
Mol Hum Reprod. 1999 Mar;5(3):214-9. doi: 10.1093/molehr/5.3.214.
The acrosome reaction (AR), an essential step for achieving mammalian fertilization, was recently introduced as a means of clinical evaluation of male fertility. However, most of the available techniques for acrosomal status assessment (except those employing electron microscopy) do not define whether the measurements represent partial or complete AR. We, therefore, performed a crossover investigation of the types of inducers and probes required for detecting partial or complete AR in human spermatozoa. The acrosomal status before and after stimulation with four AR inducers was evaluated after incubation for 3 h in capacitating conditions. We used a fluorescence-activated cell sorter with fluorescein isothiocyanate-conjugated monoclonal antibody CD46 (FITC-CD46) targeting the inner acrosomal membrane for detecting a complete AR, and fluorescein isothiocyanate-Pisum sativum agglutinin (FITC-PSA) targeting the acrosomal content for detection of both partial and complete AR. Without stimulation or following stimulation with progesterone, follicular fluid (FF) or phorbol myristate ester (PMA), the AR could be detected with FITC-PSA but not with FITC-CD46. Following stimulation with the calcium ionophore A23187, the AR could be detected by both FITC-PSA and FITC-CD46. These results suggest that spontaneous AR as well as AR induced by progesterone, PMA and FF are partial. In contrast, the AR induced by A23187 is total, i.e. both partial and complete. These findings are valuable for both research and clinical purposes and are a step towards an international agreement on a standard test for human sperm AR, for which there is an urgent need.
顶体反应（AR）是实现哺乳动物受精的关键步骤，最近被引入作为男性生育能力临床评估的一种手段。然而，大多数现有的顶体状态评估技术（电子显微镜技术除外）并未明确所测结果代表的是部分顶体反应还是完全顶体反应。因此，我们进行了一项交叉研究，以确定检测人类精子部分或完全顶体反应所需的诱导剂和探针类型。在获能条件下孵育3小时后，评估了用四种顶体反应诱导剂刺激前后的顶体状态。我们使用荧光激活细胞分选仪，用靶向顶体内膜的异硫氰酸荧光素偶联单克隆抗体CD46（FITC-CD46）检测完全顶体反应，并用靶向顶体内容物的异硫氰酸荧光素-豌豆凝集素（FITC-PSA）检测部分和完全顶体反应。在未刺激或用孕酮、卵泡液（FF）或佛波酯（PMA）刺激后，FITC-PSA可检测到顶体反应，但FITC-CD46不能。在用钙离子载体A23187刺激后，FITC-PSA和FITC-CD46均可检测到顶体反应。这些结果表明，自发顶体反应以及由孕酮、PMA和FF诱导的顶体反应是部分性的。相比之下，由A23187诱导的顶体反应是完全性的，即包括部分和完全反应。这些发现对研究和临床目的都有价值，是朝着就急需的人类精子顶体反应标准检测达成国际协议迈出的一步。
