1. We compared the ability of rat amylin, rat calcitonin gene-related peptide (CGRP) and rat and salmon calcitonins to elevate cyclic AMP levels and to inhibit [U-14C]-glucose incorporation into glycogen in insulin-stimulated intact rat soleus muscle and in two cell lines derived from rodent skeletal muscle, L6 and C2C12. 2. In intact soleus muscle, both amylin (EC50S of 0.7-6.1 nM) and salmon calcitonin (EC50S of 0.5-1.4 nM) were more potent than CGRP (EC50S of 5.6-15.8 nM) and were much more potent than rat calcitonin (EC50S of 50-137 nM) at stimulating cyclic AMP production, activating glycogen phosphorylase and inhibiting insulin-stimulated [14C]-glycogen formation. 3. In contrast, in both L6 and C2C12 cells, CGRP (EC50S of 0.042-0.12 nM) stimulated cyclic AMP formation and inhibited insulin-stimulated [U-14C]-glucose incorporation into glycogen approximately 1000 times more potently than amylin (EC50S 34-240 nM), while salmon calcitonin was without measurable effect. 4. There was a correlation between elevation of cyclic AMP and inhibition of insulin-stimulated [U-14C]-glucose incorporation into glycogen evoked by these peptides in both intact muscle (r2 = 0.69, P < 0.0004) and muscle cell lines (r2 = 0.96, P < 0.0001). 5. In conclusion, the effects of amylin, CGRP, and calcitonin on soleus muscle glycogen metabolism appear to be mediated by adenylyl cyclase-coupled receptors which show a pharmacological profile similar to high affinity amylin binding sites that have been previously reported in rat brain. In contrast, the effects of amylin and CGRP in L6 and C2C12 rodent muscle cell lines appear to be mediated by adenylyl cyclase-coupled receptors that behave like CGRP receptors.