Role of cytosolic Ca2+ in inhibition of InsP3-evoked Ca2+ release in Xenopus oocytes.

Abstract

1. Calcium liberation induced in Xenopus oocytes by flash photorelease of inositol 1,4,5-trisphosphate (InsP3) from a caged precursor was monitored by confocal microfluorimetry. The object was to determine whether inhibition of Ca2+ release seen with paired flashes arose as a direct consequence of elevated cytosolic free [Ca2+]. 2. Responses evoked by just-suprathreshold test flashes were not inhibited by subthreshold conditioning flashes, but were strongly suppressed when conditioning flashes were raised above threshold. 3. Inhibition at first increased progressively as the inter-flash interval was lengthened to about 2 s and thereafter declined, with a half-recovery at about 4 s. 4. Intracellular injections of Ca2+ caused relatively slight inhibition of InsP3-evoked signals, even when cytosolic free [Ca2+] was elevated to levels similar to those at which strong inhibition was seen in paired-flash experiments. 5. Recovery from inhibition was not appreciably slowed when Ca2+ was injected to raise the free Ca2+ level between paired flashes. 6. We conclude that inhibition of InsP3-evoked Ca2+ liberation is not directly proportional to cytosolic free Ca2+ level and that recovery from inhibition in paired-pulse experiments involves factors other than the decline of cytosolic [Ca2+] following a conditioning response.