Amylin and epinephrine did not significantly affect insulin stimulated, or basal, 3-O-methylglucose transport in isolated rat soleus muscle, as measured by the release of 3-O-methylglucose from pre-loaded tissue. Both amylin and epinephrine inhibited insulin-stimulated 2-deoxyglucose uptake (by 25% and 38%, respectively) in soleus muscle from fed rats but not from fasted rats. The latter results are consistent with amylin and epinephrine stimulating glycogenolysis and inhibiting hexokinase activity by intracellular accumulation of glucose 6-phosphate. We conclude that amylin, like epinephrine, does not specifically inhibit glucose transporters in skeletal muscle.