Steinhardt R A, Epel D
Proc Natl Acad Sci U S A. 1974 May;71(5):1915-9. doi: 10.1073/pnas.71.5.1915.
Micromolar amounts of the divalent ionophore A23187 can activate echinoderm eggs. The activations by ionophore A23187 were examined in terms of membrane elevation, the program of membrane conductance changes, the respiratory burst, and the increases in protein and DNA synthesis which normally accompany activation by sperm. In all these respects activation by the ionophore was fairly normal although subsequent cleavage and embryonic development was limited. Ionophore A23187 activations of the cortex of Lytechinus pictus and Strongylocentrotus purpuratus eggs were compared in various ionic media and were found to be completely independent of the ionic composition of the external solution. Respiration and protein synthesis of L. pictus eggs in singly substituted ionic media also indicated that these activations were independent of external sodium, calcium, or magnesium. These results suggest that the ionophore acts by releasing intracellular Ca(++). Consistent with this interpretation is the finding that eggs preloaded with (45)Ca show a 20-fold increase in (45)Ca-efflux when activated by ionophore A23187 or fertilization. Measurements of the "free" and "bound" calcium and magnesium in homogenates of the unfertilized eggs show that most of the Mg(++) is already available in the soluble form, whereas Ca(++) is sequestered but available for release. We propose that both normal fertilization and ionophore activation affect the metabolism of the egg by releasing Ca(++) sequestered in intracellular stores.
微摩尔量的二价离子载体A23187可激活棘皮动物卵。从膜隆起、膜电导变化程序、呼吸爆发以及通常伴随精子激活的蛋白质和DNA合成增加等方面研究了离子载体A23187的激活作用。尽管随后的卵裂和胚胎发育受到限制,但在所有这些方面,离子载体的激活作用相当正常。在不同离子介质中比较了离子载体A23187对花刺参和紫球海胆卵皮层的激活作用,发现其完全独立于外部溶液的离子组成。在单取代离子介质中,花刺参卵的呼吸和蛋白质合成也表明这些激活作用独立于外部的钠、钙或镁。这些结果表明,离子载体通过释放细胞内的Ca(++)起作用。与此解释一致的是,发现预先用(45)Ca加载的卵在被离子载体A23187激活或受精时,(45)Ca外流增加了20倍。对未受精卵匀浆中“游离”和“结合”的钙和镁的测量表明,大部分Mg(++)已经以可溶形式存在,而Ca(++)被螯合但可用于释放。我们提出,正常受精和离子载体激活都是通过释放细胞内储存中螯合的Ca(++)来影响卵的代谢。