Cytosolic inositol 1,4,5-trisphosphate dynamics during intracellular calcium oscillations in living cells

We developed genetically encoded fluorescent inositol 1,4,5-trisphosphate (IP₃) sensors that do not severely interfere with intracellular Ca²⁺ dynamics and used them to monitor the spatiotemporal dynamics of both cytosolic IP₃ and Ca²⁺ in single HeLa cells after stimulation of exogenously expressed metabotropic glutamate receptor 5a or endogenous histamine receptors. IP₃ started to increase at a relatively constant rate before the pacemaker Ca²⁺ rise, and the subsequent abrupt Ca²⁺ rise was not accompanied by any acceleration in the rate of increase in IP₃. Cytosolic [IP₃] did not return to its basal level during the intervals between Ca²⁺ spikes, and IP₃ gradually accumulated in the cytosol with a little or no fluctuations during cytosolic Ca²⁺ oscillations. These results indicate that the Ca²⁺-induced regenerative IP₃ production is not a driving force of the upstroke of Ca²⁺ spikes and that the apparent IP₃ sensitivity for Ca²⁺ spike generation progressively decreases during Ca²⁺ oscillations.