Arginine8-vasopressin (AVP) acts via V1 receptors (blood vessels, liver and brain) and V2 receptors (renal collecting duct). To study brain and kidney V1 receptors selectively, a specific V1 receptor antagonist [d(CH2)5,Sar7]AVP was radio-iodinated and purified by high performance liquid chromatography. Iodine-125[d(CH2)5,Sar7]AVP bound to single classes of rat liver and kidney V1 receptors with high affinity (liver: Kd = 3.0 +/- 0.9 mol/l and Bmax = 530 +/- 10 fmol/mg protein; kidney: Kd = 0.5 +/- 0.9 nmol/l and Bmax = 11 +/- 8 fmol/mg protein) in a time-dependent and saturable manner. Displacement of the radioligand from liver and renal medulla membranes and sections of the brain and kidneys by unlabelled AVP analogues was consistent with that expected for binding to V1 receptors. In vitro autoradiography of rat brain revealed areas of specific receptor binding in many regions, including regions involved in central cardiovascular regulation, such as the nucleus of the solitary tract and area postrema, as well as choroid plexus and large blood vessels. Binding was observed in several regions not previously observed to contain AVP receptors. In the kidney [3H]AVP bound to the inner and outer medulla, probably to vascular V1 and collecting duct V2 receptors. In contrast, [125I][d(CH2)5,Sar7]AVP binding was only in the inner medulla, possibly to vasa recta. These findings support a functional role for V1 receptors in the brain and kidney.