Charge movement and calcium transient were measured simultaneously in stretched frog cut twitch fibers under voltage clamp, with the internal solution containing 20 mM EGTA plus added calcium and antipyrylazo III. When the nominal free [Ca2+]i was 10 nM, the shape of the broad I(gamma) hump in the ON segments of charge movement traces remained invariant when the calcium release rate was greatly diminished. When the nominal free [Ca2+]i was 50 nM, which was close to the physiological level, the I(gamma) humps were accelerated and a slow calcium-dependent I(delta) component (or state) was generated. The peak of ON I(delta) synchronized perfectly with the peak of the calcium release rate whereas the slow decay of ON I(delta) followed the same time course as the decay of calcium release rate. Suppression of calcium release by TMB-8 reduced the amount of Q(delta) concomitantly but not completely, and the effects were partially reversible. The same simultaneous suppression effects were achieved by depleting the sarcoplasmic reticulum calcium store with repetitive stimulation. The results suggest that the mobility of Q(delta) needs to be primed by a physiological level of resting myoplasmic Ca2+. Once the priming is completed, more I(delta) is mobilized by the released Ca2+ during depolarization.