Latencies of calcium elevation and depolarization in Limulus ventral photoreceptors injected with GDP-beta S.

We have used confocal fluorescence microscopy and fluorescent calcium indicators to investigate the relationship between light-induced elevation of intracellular calcium ion concentration (Cai) and depolarization in small volumes of cytosol close to the microvillar plasma membrane of the ventral photoreceptor of Limulus polyphemus. We prolonged the latency of the light response by treatment of cells with hydroxylamine and injection of the G-protein blocker, guanosine-5'-O-(2-thiodiphosphate (GDP-beta S). Such treatment increased the latency of the cell's response from approximately 20 to 50 ms. In both treated and untreated cells we observed a close correlation between the times at which we first detected the electrical response and the elevation of Cai. We obtained 18 out of 54 and 12 out of 22 recordings, in untreated and treated cells respectively, for which the elevation of Cai was detected simultaneous with, or 1-4 ms prior to, the electrical response to light. The prolonged latent period exhibited by treated cells may make possible future investigation of the effects on the initial response to light of local photolytic release of caged compounds at the microvillar membrane.