Activation of sea-urchin eggs by a calcium ionophore

Abstract

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.