Gilkey J C, Jaffe L F, Ridgway E B, Reynolds G T
Biology Department, Purdue University, West Lafayette, Indiana 47907, USA.
J Cell Biol. 1978 Feb;76(2):448-66. doi: 10.1083/jcb.76.2.448.
Aequorin-injected eggs of the medaka (a fresh water fish) show an explosive rise in free calcium during fertilization, which is followed by a slow return to the resting level. Image intensification techniques now show a spreading wave of high free calcium during fertilization. The wave starts at the animal pole (where the sperm enters) and then traverses the egg as a shallow, roughly 20 degrees-wide band which vanishes at the antipode some minutes later. The peak free calcium concentration within this moving band is estimated to be about 30 microM (perhaps 100-1,000 times the resting level). Eggs activated by ionophore A23187 may show multiple initiation sites. The resulting multiple waves never spread through each other; rather, they fuse upon meeting so as to form spreading waves of compound origin. The fertilization wave is nearly independent of extracellular calcium because it is only slightly slowed (by perhaps 15%) in a medium containing 5 mM ethylene glycol-bis[beta-aminoethyl ether]N,N'-tetraacetic acid (EGTA) and no deliberately added calcium. It is also independent of the large cortical vesicles, which may be centrifugally displaced. Normally, however, it distinctly precedes the well-known wave of cortical vesicle exocytosis. We conclude that the fertilization wave in the medaka egg is propagated by calcium-stimulated calcium release, primarily from some internal sources other than the large cortical vesicles. A comparison of the characteristics of the exocytotic wave in the medaka with that in other eggs, particularly in echinoderm eggs, suggests that such a propagated calcium wave is a general feature of egg activation.
将水母发光蛋白注入青鳉(一种淡水鱼)的卵中,在受精过程中可观察到游离钙含量急剧上升,随后缓慢恢复到静息水平。图像增强技术显示,受精过程中会出现游离钙含量高的扩散波。该波从动物极(精子进入的位置)开始,然后以约20度宽的浅带形式穿过卵子,几分钟后在对极消失。据估计,这个移动带内的游离钙峰值浓度约为30微摩尔(可能是静息水平的100 - 1000倍)。由离子载体A23187激活的卵可能会出现多个起始位点。由此产生的多个波不会相互穿过;相反,它们相遇时会融合,从而形成复合起源的扩散波。受精波几乎与细胞外钙无关,因为在含有5毫摩尔乙二醇双(β - 氨基乙基醚)N,N' - 四乙酸(EGTA)且未特意添加钙的培养基中,其速度仅略有减慢(约15%)。它也与可能被离心移位的大皮质囊泡无关。然而,正常情况下,它明显先于众所周知的皮质囊泡胞吐波。我们得出结论,青鳉卵中的受精波是由钙刺激的钙释放介导的,主要来自大皮质囊泡以外的一些内部来源。将青鳉中的胞吐波特征与其他卵,特别是棘皮动物卵中的特征进行比较,表明这种传播的钙波是卵激活的一个普遍特征。