The aims of this study were to quantify the Ca2+ release underlying twitch contractions of mammalian fast- and slow-twitch muscle and to comprehensively describe the transient inactivation of Ca2+ release following a stimulus. Experiments were performed using bundles of fibres from mouse extensor digitorum longus (EDL) and soleus muscles. Ca2+ release was quantified from the amount of ATP used to remove Ca2+ from the myoplasm following stimulation. ATP turnover by crossbridges was blocked pharmacologically (N-benzyl-p-toluenesulphonamide for EDL, blebbistatin for soleus) andmuscle heat production was used as an index of Ca2+ pump ATP turnover. At 20?C, Ca2+ release in response to a single stimulus was 34 and 84 孯l/(kg muscle) for soleus and EDL, respectively, and increased with temperature (30?C: soleus, 61 孯l/kg; EDL, 168 孯l/kg). Delivery of another stimulus within 100 ms of the first produced a smaller Ca2+ release. The maximum magnitude of the decrease in Ca2+ release was greater inEDLthan soleus. Ca2+ release recovered with an exponential time course which was faster in EDL (mean time constant at 20?C, 32.1 ms) than soleus (65.6 ms) and faster at 30?C than at 20?C. The amounts of Ca2+ released and crossbridge cycles performed are consistent with a scheme in which Ca2+ binding to troponin-C allowed an average of ~1.7 crossbridge cycles in the two muscles.