Distributional Change of Membrane-associated Calcium in Sea Urchin Eggs during Fertilization

The "source and sink" for the intracellular calcium released during fertilization were examined in sea urchin eggs, Hemicentrotus pulcherrimus , with chlortetracycline as a fluorescent chelate probe. In order to distinguish the differential distribution of membrane-associated calcium in various compartments in cytoplasm, eggs were stratified by centrifugation before or after fertilization. Only the layer containing mainly mitochondria exhibited the chlortetracycline-fluorescence in unfertilized eggs. After fertilization, a new fluorescent band emerged in the membrane-rich clear layer of stratified eggs. Chlortetracycline-fluorescence in the clear layer was gradually redistributed surrounding the prophase nucleus and then incorporated into the mitotic apparatus. From these observations, we postulate that the major source(s) of released free calcium ions at fertilization is in the mitochondira layer and membranes in the clear layer are newly activated as the calcium sequestering system after fertilization.