Guerrero A, Darszon A
Departamento de Bioquímica, Centro de Investigación y de Estudios Avanzados, México City, Mexico.
Biochim Biophys Acta. 1989 Mar 27;980(1):109-16. doi: 10.1016/0005-2736(89)90206-x.
Sea urchin sperm must undergo the acrosome reaction to fertilize eggs. The natural inducer of this reaction is the most external coat of the egg, named 'jelly'. The ionic composition of the extracellular and intracellular media and the permeability properties of the sperm plasma membrane are fundamental in this reaction. As Ca2+ is required for the acrosome reaction to occur, its intracellular concentration ([Ca2+]i) was measured with fura-2. In 10 mM Ca2+, egg jelly induced the acrosome reaction and an increase in [Ca2+]i that lasted for several minutes. However, at 0.5 or 2 mM Ca2+, it became evident that the Ca2+-influx pathway activated by jelly opened only for a few seconds; this prevented both the full increase in [Ca2+]i and the acrosome reaction even after the concentration of Ca2+ was raised to 10 mM. In the presence of jelly, the time this permeability pathway remained open was inversely related to the extracellular concentration of Ca2+ ([ Ca2+]e). Using Bisoxonol (a permeant fluorescent membrane potential probe), it was found that the jelly-induced depolarization depended on [Ca2+]e and was proportional to the increase in [Ca2+]i. Since [Ca2+]i could affect the jelly-induced Ca2+ influx through calmodulin, two of its antagonists, trifluoperazine and W-7, were tested. Both compounds blocked the acrosome reaction by inhibiting the jelly-induced increase in [Ca2+]i. W-5 at the same concentration had no effect. The results suggest that one of the jelly-activated Ca2+-influx pathways, probably a channel, is the target of the calmodulin antagonists.
海胆精子必须经历顶体反应才能使卵子受精。这种反应的天然诱导剂是卵子最外层的包膜,称为“卵胶”。细胞外和细胞内介质的离子组成以及精子质膜的通透性特性在这一反应中至关重要。由于顶体反应的发生需要Ca2+,因此用fura-2测量了其细胞内浓度([Ca2+]i)。在10 mM Ca2+条件下,卵胶诱导顶体反应并使[Ca2+]i增加,这种增加持续了几分钟。然而,在0.5 mM或2 mM Ca2+条件下,很明显卵胶激活的Ca2+内流途径仅开放了几秒钟;这就阻止了[Ca2+]i的充分增加以及顶体反应,即使将Ca2+浓度提高到10 mM也是如此。在有卵胶存在的情况下,这种通透性途径保持开放的时间与细胞外Ca2+浓度([Ca2+]e)呈负相关。使用双苯甲酰羟肟酸(一种可渗透的荧光膜电位探针)发现,卵胶诱导的去极化取决于[Ca2+]e,并且与[Ca2+]i的增加成正比。由于[Ca2+]i可以通过钙调蛋白影响卵胶诱导的Ca2+内流,因此测试了它的两种拮抗剂三氟拉嗪和W-7。这两种化合物都通过抑制卵胶诱导的[Ca2+]i增加来阻断顶体反应。相同浓度的W-5没有作用。结果表明,卵胶激活的Ca2+内流途径之一,可能是一个通道,是钙调蛋白拮抗剂的作用靶点。