Ca2+i transients and Ca2+i‐dependent chloride current in single Purkinje cells from rabbit heart.

Single Purkinje cells, enzymatically isolated from rabbit ventricle, were studied under whole-cell voltage clamp and internally perfused with the fluorescent Ca2+ indicator, indo-1 (100 microM). 2. Fast Ca2+i transients were elicited by brief depolarizations from a holding voltage of -45 mV and by repolarization from very positive potentials. The peak Ca2+i-voltage relation was bell-shaped with a peak around +10 mV. 3. Ca2+i transients were completely blocked by the Ca2+ channel antagonist, nisoldipine (10 microM) and were very small when Ca2+ release from the sarcoplasmic reticulum (SR) was prevented by superfusion of cells by caffeine (1 mM) or ryanodine (10 microM). A fast application of caffeine induced a transient increase in Ca2+i. These results suggest Ca2+i transients are due to Ca(2+)-induced Ca2+ release from the SR. 4. Rate of decline of the Ca2+i transient was voltage dependent, suggesting contribution of the Na(+)-Ca2+ exchanger to Ca2+ efflux. At very positive potentials (> +60 mV), Ca2+ influx through the Na(+)-Ca2+ exchanger could be observed. 5. A transient outward current was observed at potentials positive to +10 mV, but only if depolarizing pulses were accompanied by a Ca2+i transient. 6. When the amplitude of the Ca2+i transient was changed by (1) changes in Ca2+o, (2) changes in frequency of depolarization or (3) conditioning prepulses, the amplitude of the outward current changed in the same direction. This suggests activation of the current is dependent on and graded by Ca2+i. 7. The outward current was observed in K(+)-free solutions, in the presence of Cs+ and TEA+, and was not blocked by 4-aminopyridine (10 mM). In contrast, DIDS (100 microM) decreased the outward current by 70 +/- 20% (mean +/- S.D., n = 9), without affecting Ca2+i. 8. When external Cl- was lowered, the amplitude of the outward current decreased; when internal Cl- was replaced by aspartate, it became apparent at more negative potentials. These interventions strongly suggest the current was carried by Cl-; it can therefore be referred to as a Ca2+i-activated Cl- current or ICl(Ca). 9. When ICl(Ca) was maximally activated during a conditioning step, steps to negative potentials revealed inward currents through ICl(Ca) (in symmetrical Cl- solutions). The fully activated I-V relation was linear. 10. ICl(Ca) could be activated at membrane potentials between -80 and +80 mV by a fast application of caffeine (10 mM), inducing Ca2+ release from the SR, demonstrating that ICl(Ca) does not require membrane depolarization or Ca2+ influx through the Ca2+ channel for its activation.(ABSTRACT TRUNCATED AT 400 WORDS)