The tridecapeptide neurotensin (NT) contracts the guinea pig ileum through a neurogenic process that is mediated in part by acetylcholine and substance P and relaxes the guinea pig colon through a direct action on smooth muscle cells involving the opening of Ca(++)-dependent K+ channels. The non-peptide NT antagonist, SR 48692 (2-[1-(7-chloro-4-quinolinyl)-5-(2,6- dimethoxyphenyl)pyrazol-3-yl)carbonylamino]tricyclo-(3.3.1.1 .3.7)decan-2- carboxylic acid), potently inhibited NT binding to membranes prepared from the guinea pig ileum and colon with Ki values of approximately 3 nM. SR 48527 ((S)-(+)-[1-(7-chloro-4-quinolinyl)-5-(2,6-dimethoxyphenyl)pyrazol-3- yl)carbonylamino]cyclohexylacetic acid) and SR 49711 ((R)-(-)-[1-(7-chloro-4-quinolinyl)-5-(2,6-dimethoxyphenyl)pyrazol- 3-yl)carbonylamino]cyclohexylacetic acid), two enantiomers structurally related to SR 48692, were respectively equipotent with and a 100-fold less potent than SR 48692 in inhibiting NT binding in both tissues. In both membrane preparations, NT binding was increased by Mg++ and decreased by Na+ and guanosine 5'-[gamma-thio]triphosphate, whereas SR 48692 binding was not significantly affected by these agents. SR 48692 inhibited NT-induced contraction and relaxation in guinea pig ileum and colon preparations, respectively, with Ki values between 4 and 5 nM. As in binding studies, SR 48527 was as potent, whereas SR 49711 was 100-fold less potent than SR 48692 in antagonizing NT responses in both the guinea pig ileum and colon. Altogether, our results show that NT receptors in the guinea pig ileum and colon, although functionally distinct, are coupled to G-proteins and display similar biochemical and pharmacological properties, in particular with regard to their sensitivity and stereoselectivity toward nonpeptide antagonists related to SR 48692. Because of their high potency to antagonize NT actions in intestinal preparations, SR 48692 and SR 48527 represent useful tools to study the physiological role of NT in the digestive tract.