Calcitonin gene-related peptide (CGRP), a 37-amino acid peptide, is widely distributed in the gastrointestinal tract where it is colocalized with substance P. The effect of CGRP on gallbladder motility is unknown. The objective of this study was to examine the effect of CGRP on cholecystokinin-octapeptide (CCK-8) and substance P-stimulated gallbladder contraction in vivo and in vitro. In in vivo studies intragallbladder pressure was measured in response to bolus administration of CCK-8 (10(-15) to 10(-9) mol/kg) or substance P (10(-12) to 10(-7) mol/kg), either alone or with a continuous infusion of CGRP (10(-9) mol/kg/hr), in anesthetized guinea pigs. In in vitro studies the contractile force of guinea pig gallbladder muscle strips was examined in response to CCK-8 (10(-12) to 10(-7) mol/L) and substance P (10(-9) to 10(-6) mol/L), alone or with CGRP (10(-10) to 10(-6) mol/L). CGRP (10(-9) mol/kg/hr) inhibited in vivo gallbladder contraction that was stimulated by CCK-8, but not by substance P. CGRP alone produced a significant (p less than 0.05) dose-related decrease in the resting tension of gallbladder strips in vitro. CGRP (10(-6) mol/L) inhibited gallbladder muscle tension in vitro, stimulated by both CCK-8 and substance P. These studies show that CGRP can affect gallbladder motor activity by decreasing smooth muscle tone and that CGRP can antagonize the action of CCK and substance P. CGRP may be involved in the physiologic control of gallbladder emptying and refilling.