1. Potency orders were determined for a series of agonists and antagonists on the calcitonin gene-related peptide (CGRP) receptor of rat L6 myocytes. The agents tested were all shown to have been active against CGRP, amylin or adrenomedullin receptors. 2. AC187 had a pIC50 of 6.8 +/- 0.10, making it 14 fold less potent as an antagonist than CGRP8-37 (pIC50, 7.95 +/- 0.14). Amyline8-37 was equipotent to AC187 (pIC50, 6.6 +/- 0.16) and CGRP19-32 was 3 fold less potent than either (pIC50, 6.1 +/- 0.24). 3. [Ala11]-CGRP8-37 was 6 fold less potent than CGRP8-37, (pIC50, 7.13 +/- 0.14), whereas [Ala18]-CGRP8-37 was approximately equipotent to CGRP8-37 (pIC50, 7.52 +/- 0.15). However, [Ala11,Ala18]-CGRP8-37 was over 300 fold less potent than CGRP8-37 (pIC50, 5.30 +/- 0.04). 4. [Tyr0]-CGRP28-37, amylin19-37 and adrenomedullin22-52 were inactive as antagonists at concentrations of up to 1 microM. 5. Biotinyl-human alpha-CGRP was 150 fold less potent than human alpha-CGRP itself (EC50 values of 48 +/- 17 nM and 0.31 +/- 0.13 nM, respectively). At 1 microM, [Cys(acetomethoxy)2,7]-CGRP was inactive as an agonist. 6. These results confirm a role for Arg11 in maintaining the high affinity binding of CGRP8-37. Arg18 is of less direct significance for high affinity binding, but it may be important in maintaining the amphipathic nature of CGRP and its analogues.