Sarcolemmal membranes were isolated from skeletal muscle by a sucrose density gradient method from rats with diabetes induced by a streptozotocin injection (65 mg/kg iv). The activities of Na(+)-dependent Ca2+ uptake and Ca2(+)-stimulated adenosine-triphosphatase (ATPase) in the sarcolemmal fraction from diabetic rats was higher than those from the control animals. These changes were apparent at various times of incubation (1-10 min) as well as at different concentrations of free Ca2+ (10(-7) to 10(-5) M) and developed during the third and/or fourth weeks after streptozotocin injection. ATP-dependent Ca2+ uptake in the sarcolemmal vesicles was also increased at 28 and 56 days after inducing diabetes. Treatment of diabetic animals with insulin for 14 days reversed the changes in Ca2+ transport activities toward the control levels. Sarcolemmal Mg2(+)-ATPase and Na(+)-K(+)-ATPase activities remained unchanged in diabetic preparations. Furthermore, no difference in the sarcolemmal phospholipid composition and sodium dodecyl sulfate-gel electrophoretic pattern was evident between the control and experimental groups. These results indicate a higher activity of the sarcolemmal Ca2+ transport, which may be associated with hyperfunction of the skeletal muscle in diabetic rats.