In functionally skinned muscle fibers from the rabbit, we studied the effect of propranolol on calcium activation of the contractile proteins and, in separate experiments, on calcium uptake and release from the sarcoplasmic reticulum (SR) while measuring physiological tension. Pieces from isolated papillary muscle (PM), soleus (SL) (slow-twitch skeletal muscle), and adductor magnus (AM) (fast-twitch skeletal muscle) were homogenized (sarcolemma disrupted). A fiber bundle from PM and single fibers from SL and AM were dissected from the homogenate and mounted on a photodiode tension transducer. To study Ca2+-activated tension development of the contractile proteins, we used high EGTA (7 mmol/l) to control the free calcium concentration. To study SR function, we used five different solutions to load the calcium into the SR and to release it from the SR with 25 mmol/l caffeine, thus producing a tension transient. In general, propranolol has similar mechanisms of action in the three muscle types. Propranolol (0.1-1.0 mmol/l) increased the submaximal calcium-activated tension development in all muscles but with PM = SL greater than AM, and this increase was correlated with increases in calcium binding to isolated troponin C. Propranolol increased the maximal calcium-activated tension development in PM and SL, but decreased that in AM. Propranolol at concentrations of 0.3-1.0 mmol/l decreased calcium uptake by the SR but did not change calcium release in any of the three muscles. In PM, however, propranolol at a concentration of 0.1 mmol/l increased calcium uptake by the SR. We conclude that propranolol induces decreases in muscle contraction mainly by decreasing calcium uptake by the SR.