Assessment of acrosomal status in rat spermatozoa: studies on carbohydrate and non-carbohydrate agonists
Assessment of acrosomal status in rat spermatozoa: studies on carbohydrate and non-carbohydrate agonists
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DOI：
10.1016/s0003-9861(02)00278-3
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发表时间：
2002-08-01
影响因子：
3.9
通讯作者：
Tulsiani, DRP
中科院分区：
文献类型：
--
作者：
Bendahmane, M;Zeng, HT;Tulsiani, DRP
在小鼠和其他几个物种中，包括人，获能的顶体完整精子与天然[可溶性透明质酸（ZP）和孕酮（P4）]和合成[新糖蛋白（ngps）和钙（Ca2+）离子载体]激动剂相互作用，然后启动Ca2+依赖性信号转导级联反应。最终结果是精子质膜在多个位点与顶体外膜融合，顶体内容物胞吐[即，顶体反应（AR）。这一步骤被认为是使顶体反应的精子能够穿透ZP并使卵子受精的先决条件。虽然大鼠是最常用的实验室动物之一，但对该种属中诱导AR的激动剂的化学性质知之甚少。缺乏这些信息主要是由于大鼠精子顶体是一个相对较薄的结构。因此，很难评估该物种精子顶体的状态。在这份报告中，我们描述了使用考马斯亮蓝染色程序，以评估的状态，大鼠精子顶体的光学显微镜。该程序是高度可重复的，使我们能够确定碳水化合物（NGPS和小鼠ZP）和非碳水化合物（P4和Ca2+离子载体）激动剂对获能精子的影响。此外，我们已经使用了药理学的方法来检查钙调素（CaM），钙离子结合蛋白，在精子中的AR诱导的功能意义。本报告中提供的数据表明，几种ngps，增溶的mZP，P4和Ca2+离子载体诱导大鼠精子中的AR。此外，我们证明，而钙调素拮抗剂阻断P4诱导的AR，大多数使用的抑制剂对钙离子载体诱导的（非生理性）AR没有显着影响。(C)2002 Elsevier Science（美国）。All rights reserved.
In the mouse and several other species, including man, capacitated acrosome-intact spermatozoa interact with natural [soluble zona pellucida (ZP) and progesterone (P4)] and synthetic [neoglycoproteins (ngps) and calcium (Ca2+) ionophore] agonists, prior to the initiation of a Ca2+-dependent signal transduction cascade. The net result is the fusion of the sperm plasma membrane overlying the outer acrosomal membrane at multiple sites and exocytosis of acrosomal contents [i.e., induction of the acrosome reaction (AR)]. This step is believed to be a prerequisite that enables the acrosome-reacted spermatozoon to penetrate the ZP and fertilize the egg. Although the rat is one of the most commonly used laboratory animals, very little is known about the chemical nature of agonists that induce the AR in this species. The lack of this information is primarily due to the fact that the rat sperm acrosome is a relatively thin structure. Thus, it is difficult to assess the status of the sperm acrosome in this species. In this report, we describe the use of a Coomassie brilliant blue dye staining procedure to assess the status of the rat sperm acrosome by light microscopy. The procedure is highly reproducible and has allowed us to determine the effects of carbohydrate (ngps and mouse ZP) and noncarbohydrate (P4 and Ca2+ ionophore) agonists on capacitated spermatozoa. In addition, we have used a pharmacological approach to examine the functional significance of calmodulin (CaM), a Ca2+ -binding protein, in induction of the AR in spermatozoa. Data presented in this report demonstrate that several ngps, solubilized mZP, P4, and Ca2+ ionophores induce the AR in rat spermatozoa. Furthermore, we demonstrate that, whereas CaM antagonists blocked P4-induced AR, most of the inhibitors used had no significant effect on the Ca2+ ionophore-induced (nonphysiological) AR. (C) 2002 Elsevier Science (USA). All rights reserved.