The role of Ca influx in cardiac muscle excitation-contraction coupling. Assessment by extracellular Ca microelectrodes.

Tension, dT/dt, and extracellular free [Ca] [( Ca]0) were continuously measured in isolated rabbit right papillary muscles at 28 degrees C. Double-barreled Ca-selective microelectrodes incorporating Simon's neutral Ca exchange resin (ETH-1001) were used to monitor [Ca]0. Decreases of [Ca]0 were seen during the course of single twitches, before the development of significant tension. This depletion of Ca0 probably represents Ca influx into the cells from the extracellular space. The magnitude of this Ca0 depletion is decreased by Co, verapamil, long rest intervals, and reduction of [Ca]0. The magnitude is increased by catecholamines, reduction of [Na]0, caffeine, continued pacing, and elevation of [Ca]0. After 3-min rest, stimulation (0.5-1 Hz) produces a biphasic tension response (larger first beat, small second beat, and monotonic rise to control). Caffeine (5 mM) changes the pattern after rest to a monotonic increase. Ca influx increases monotonically in both cases. Addition of 20 mM Co during the rest reduces tension of all beats by similar amounts in the presence of caffeine. In the absence of caffeine, Co has a much weaker effect on the first beat than on subsequent beats. The results suggest that caffeine inhibits an intracellular component of activator Ca that is more important after a rest interval, but that Ca influx becomes increasingly more important during continued pacing under the conditions used here.