Article,

Effects of partial sarcoplasmic reticulum calcium depletion on calcium release in frog cut muscle fibers equilibrated with 20 mM EGTA.

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J. Gen. Physiol., 112 (3): 263--295 (September 1998)

Abstract

Resting sarcoplasmic reticulum (SR) Ca content (CaSRR) was varied in cut fibers equilibrated with an internal solution that contained 20 mM EGTA and 0-1.76 mM Ca. SR Ca release and CaSRR were measured with the EGTA-phenol red method (. J. Gen. Physiol. 106:259-336). After an action potential, the fractional amount of Ca released from the SR increased from 0.17 to 0.50 when CaSRR was reduced from 1, 200 to 140 microM. This increase was associated with a prolongation of release (final time constant, from 1-2 to 10-15 ms) and of the action potential (by 1-2 ms). Similar changes in release were observed with brief stimulations to -20 mV in voltage-clamped fibers, in which charge movement (Qcm) could be measured. The peak values of Qcm and the fractional rate of SR Ca release, as well as their ON time courses, were little affected by reducing CaSRR from 1,200 to 140 microM. After repolarization, however, the OFF time courses of Qcm and the rate of SR Ca release were slowed by factors of 1.5-1.7 and 6.5, respectively. These and other results suggest that, after action potential stimulation of fibers in normal physiological condition, the increase in myoplasmic free Ca that accompanies SR Ca release exerts three negative feedback effects that tend to reduce additional release: (a) the action potential is shortened by current through Ca-activated potassium channels in the surface and/or tubular membranes; (b) the OFF kinetics of Qcm is accelerated; and (c) Ca inactivation of Ca release is increased. Some of these effects of Ca on an SR Ca channel or its voltage sensor appear to be regulated by the value of Ca within 22 nm of the mouth of the channel.

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