Effect of diazepam on calcium translocation during physiological muscle fatigue

Stimulation of frog sartorius muscle at 1 Hz leads to an initial positive staircase during the first 120 twitches and is followed by a negative staircase. There is a net calcium influx into two distinct compartments within the muscle during the positive staircase. The two compartments are separated by measuring the calcium extracted from muscles soaked in strontium-Ringer for 15 min and the calcium remaining in the muscle. A net gain of extractable Ca++ (0.32 mumol/g wet wt.) and residual Ca++ (0.18 mumol/g) is observed during positive staircase. A loss in residual Ca++, a gain in extractable Ca++ and a net loss of Ca++ (0.09 mumol/g) to the bathing medium occur during the period preceding physiological muscle fatigue (60 to 120 twitches). Diazepam (EC50, 5.6 X 10(-6) M) causes a marked reduction in the latent period and increases the rate constant 2.6 times the control value for physiological muscle fatigue. A net loss of 0.31 mumol/g of Ca++ to the bathing medium occurs during the interval between 60 and 120 twitches. Diazepam increases net Ca++ efflux 3.5-fold during this interval when compared to control muscles. Diazepam does not affect the Ca++ gained during the positive staircase but accelerates the loss of calcium from the residual and the extractable compartments during the initial phase of physiological muscle fatigue. Physiological muscle fatigue is attributed to an accumulation of calcium in the transverse tubular network and an uncoupling of the muscle action potential from contraction.