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The decay of the Ca2+ current (ICa) during a maintained depolarization was studied in intact twitch skeletal muscle fibres of Rana pipiens and Rana moctezuma with the three-micro-electrode voltage-clamp technique. ICa was recorded at 23 degrees C, after blocking K+ currents, in TEA methanesulphonate saline with 10 mM-Ca2+ made hypertonic by adding 350 mM-sucrose. In two-pulse experiments, ICa during the test pulse was reduced to about 80% (R. pipiens) or 50% (R. moctezuma) of the control value, without any detectable inward ICa during 7 s conditioning pre-pulses. The experimental points of the steady-state inactivation curve (h infinity) were fitted to h infinity = (1 + exp [Em - Vh)/kh]-1, where Em is the membrane potential and with Vh = -33 +/- 3 mV and kh = 6 +/- 1 mV for R. pipiens, and Vh = -44 +/- 3 mV and kh = 9.5 +/- 1.0 mV for R. moctezuma. The rate constant of decay for inactivated currents (range -8 to -47 mA cm-3) and for control currents (range -23 to -62 mA cm-3), was independent of ICa amplitude. The average rate constant of decay at 0 mV was 1.18 +/- 0.02 s-1 (66). These results indicate that in intact fibres under hypertonic solution ICa decay can be explained by a voltage-dependent inactivation process and not by depletion of tubular Ca2+. The absence of depletion could be due to a large fractional tubular volume or to the presence of a Ca2+ pump in the tubular system.

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