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储存式钙通道触发海胆精子顶体囊泡的胞吐作用。

Store-operated calcium channels trigger exocytosis of the sea urchin sperm acrosomal vesicle.

作者信息

Hirohashi Noritaka, Vacquier Victor D

机构信息

Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California San Diego, La Jolla, CA 92093-0202, USA.

出版信息

Biochem Biophys Res Commun. 2003 May 2;304(2):285-92. doi: 10.1016/s0006-291x(03)00587-4.

Abstract

The acrosome reaction (AR) of sperm is a prerequisite for fusion with the egg. In sea urchins, the complete AR (CAR) consists of exocytosis of the acrosomal vesicle (AV) and polymerization of acrosomal actin to form the approximately 1 micro m long acrosomal process. The fucose sulfate polymer (FSP) of egg jelly stimulates Ca(2+) entry through two distinct Ca(2+) channels and induces the CAR. Here we report that the second channel is blocked by SKF96365 (SKF), an inhibitor of store-operated channels. SKF also blocks the thapsigargin (TG), trifluoperazine (TFP), and calmidizolium (CMZ) stimulated Ca(2+) entry into sperm. These data indicate that the second Ca(2+) channel is a store-operated channel (SOC) that may be regulated by calmodulin. The TG, TFP, and CMZ-induced intracellular Ca(2+) elevations are similar to those induced by FSP, but the sperm acrosomal process does not polymerize. An antibody to bindin, the major protein of the AV, showed that in a significant percentage of these drug-treated sperm, the AV had undergone exocytosis. When NH(4)Cl was added to increase intracellular pH, the TG-treated sperm polymerized actin to form the acrosomal process. We conclude that the second Ca(2+) channel of sea urchin sperm is a SOC that triggers AV exocytosis.

摘要

精子的顶体反应(AR)是与卵子融合的先决条件。在海胆中,完整的顶体反应(CAR)包括顶体小泡(AV)的胞吐作用以及顶体肌动蛋白聚合形成约1微米长的顶体突起。卵胶的硫酸岩藻糖聚合物(FSP)通过两个不同的Ca(2+)通道刺激Ca(2+)内流并诱导CAR。在此我们报告,第二个通道被储存操纵性通道抑制剂SKF96365(SKF)阻断。SKF还阻断毒胡萝卜素(TG)、三氟拉嗪(TFP)和平痛新(CMZ)刺激的Ca(2+)进入精子。这些数据表明,第二个Ca(2+)通道是一个可能受钙调蛋白调节的储存操纵性通道(SOC)。TG、TFP和CMZ诱导的细胞内Ca(2+)升高与FSP诱导的相似,但精子顶体突起不会聚合。针对顶体小泡主要蛋白结合素的抗体显示,在这些药物处理的精子中,有相当比例的顶体小泡已经发生了胞吐作用。当加入NH(4)Cl以提高细胞内pH值时,经TG处理的精子使肌动蛋白聚合形成顶体突起。我们得出结论,海胆精子的第二个Ca(2+)通道是一个触发顶体小泡胞吐作用的SOC。

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