Effects of caffeine on intracellular calcium concentrations in frog skeletal muscle fibres.

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1. The mechanism of twitch potentiation by caffeine was studied in single muscle fibres dissected from m. tibialis anterior of Rana temporaria. Fibres were injected with a Ca2+-sensitive photo-protein, aequorin, and the resultant light signal and tension were simultaneously recorded. 2. In twitch responses triggered every 60 s, peaks of light and tension were maintained at a constant level. Low concentrations of caffeine (0.2-0.4 mM) potentiated twitch tension accompanied by a slight increase in light signal. 3. Although peak twitch tension was dose-dependently potentiated by caffeine, light peaks were suppressed at relatively higher concentrations of caffeine (0.6-1.5 mM) with prolonged decay of the light and tension signals. 4. Light intensity in the resting muscle (resting glow) was elevated by caffeine (0.2-1.5 mM) dose-dependently without detectable tension development. This effect of caffeine was suppressed by 0.5-1.0 mM-procaine or 10 mM-adenine, inhibitors of Ca2+-induced Ca2+ release. 5. In caffeine-treated preparations, peaks of light and tension were augmented by application of procaine (0.5-1.0 mM). Adenine (10 mM) affected the light signal in essentially the same way as procaine, but the effect varied from fibre to fibre. 6. From these results, the following hypothesis is proposed: low concentrations of caffeine directly induce Ca2+ release from sarcoplasmic reticulum (s.r.) in in the resting state, and facilitates Ca2+ release from s.r. induced by the action potential during twitch. At relatively higher concentrations of caffeine (0.6-1.5 mM), rise of resting [Ca2+]i (intracellular free Ca2+ concentration) before activation might be an important factor in twitch potentiation by caffeine. If the rise of resting [Ca2+]i induced by caffeine is inhibited by procaine and the content of Ca2+ in s.r. is well maintained, caffeine could facilitate Ca2+ release by depolarization and thus potentiate twitch tension.

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