The Ca2+ permeability of rabbit skeletal muscle sarcolemmal vesicles was investigated by means of radioisotope flux measurements. A membrane vesicle fraction highly enriched in sarcolemma, as revealed by enzymatic markers, was obtained from the 22-27% region of sucrose gradients after isopycnic centrifugation. The ability of sarcolemmal vesicles to exchange Na+ for Ca2+ was investigated by measuring Ca2+ influx into and efflux from sarcolemmal vesicles in the presence and absence of a Na+ gradient. It was found that Ca2+ movements were enhanced in the direction of the higher Na+ concentration. When intra-and extravesicular Na+ concentrations were high, Na+-Na+ exchanged predominated and Na+-Ca2+ exchange was low or absent. The presence of the Ca2+ ionophore A23187 in the dilution medium resulted in the rapid release of Ca2+ and the elimination of the Na+-enhanced efflux of Ca2+, suggesting that internal rather than bound external Ca2+ was exchanged with Na+ . La3+ abolished Na+-Ca4+ exchanged was not due to sarcoplasmic reticulum or mitochondrial contaminants. This investigation suggests that skeletal muscle, like cardiac muscle and neurons, is capable of a transmembranous Na/-Ca2+ exchange.