Ca2+‐dependent inactivation of Ca2+‐induced Ca2+ release in bullfrog sympathetic neurons

We studied inactivation of Ca2+‐induced Ca2+ release (CICR) via ryanodine receptors (RyRs) in bullfrog sympathetic neurons. The rate of rise in [Ca2+]i due to CICR evoked by a depolarizing pulse decreased markedly within 10–20 ms to a much slower rate despite persistent Ca2+ entry and little depletion of Ca2+ stores. The Ca2+ entry elicited by the subsequent pulse within 50 ms, during which the [Ca2+]i level remained unchanged, did not generate a distinct [Ca2+]i rise. This mode of [Ca2+]i rise was unaffected by a mitochondrial uncoupler, carbonyl cyanide p‐trifluromethoxy‐phenylhydrazone (FCCP, 1 μm). Paired pulses of varying interval and duration revealed that recovery from inactivation became distinct ≥ 50 ms after depolarization and depended on [Ca2+]i. The inactivation was prevented by BAPTA (≥ 100 μm) but not by EGTA (≤ 10 mm), whereas the activation was less affected by BAPTA. When CICR was partially activated, some of the non‐activated RyRs were also inactivated directly. Thus, the inactivation in these neurons is induced by Ca2+ binding to the high‐affinity regulatory sites residing very close to Ca2+ channels and/or RyRs, although the sites for activation are located much closer to those Ca2+ sources. The rate of [Ca2+]i decay after the pulse decreased with increasing pulse duration longer than 10 ms, and this was abolished by BAPTA. Thus, some mechanism counteracting Ca2+ clearance is induced after full inactivation and potentiated during the pulse. Possible models for RyR inactivation were proposed and the roles of inactivation in Ca2+ signalling were discussed.