1. Shortening and intracellular Ca2+ (Ca(i)2+) transients were measured in isolated rabbit ventricular myocytes during paired contractures induced by rapid application of 10 mM caffeine 2. Caffeine-induced contractures relax despite maintained presence of caffeine. In control solution, a second phasic caffeine contracture failed to appear, unless the sarcoplasmic reticulum (SR) was refilled by a series of electrically stimulated twitches during the interval between caffeine exposures. 3. The relaxation of caffeine-induced contractures in 0 Na+-0 Ca2+ solution has previously been shown to rely on mitochondrial Ca2+ uptake and sarcolemmal Ca2+-ATPase. Thus, a second caffeine contracture (T2) while still in O Na+-0 Ca2+ was greatly reduced compared to the first one (T1). However, the amplitude of T2 increased exponentially with the time interval, attaining a maximum of ~ 50% of T1 for an interval of 180-300 s, with a time constant (Υ) of 41.2 s. Similar results were found for Ca(i)2+ transients (Υ = 45 S). 4. Inhibition of the mitochondrial Ca2+ uptake by the oxidative phosphorylation uncoupler, FCCP during T1 dramatically depressed T2. On the other hand, inhibition of the sarcolemmal Ca2+-ATPase (by increasing extracellular Ca2+ concentration, [Ca2+](o)) resulted in increase of T2. Spermine inclusion during T1 also increased T2, possibly by an increase of mitochondrial Ca2+ uptake. 5. We conclude that Ca2+ taken up by mitochondria during the decline of T1 moves back to the SR after caffeine is removed, with a τ ~40 s. 6. Partial intracellular :Na+ depletion by prolonged (3 min) perfusion with O Na+ Ca2+,solution before T1 (a) accelerated relaxation and [Ca2+](i) decline during T1, and (b) slowed, but did not abolish, the recovery of T2 as the interval was increased. This effect was particularly pronounced when choline was used instead of Li+ as the Na+ substitute. 7. We further conclude that the mitochondrial Na+-Ca2+ antiporter influences the rate of net Ca2+ uptake by mitochondria and is also important in Ca2+ efflux from mitochondria during rest.
|Original language||English (US)|
|Number of pages||19|
|Journal||Journal of Physiology|
|State||Published - 1993|
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