In vitro autoradiography of GTPgamma[35S] binding at activated NPY receptor subtypes in adult rat brain

Abstract

Guanylyl 5'-[gamma[35S]thio]-triphosphate (GTPgamma[35S]) binding to NPY receptor-activated G-proteins was measured in adult rat brain sections in order to determine the neuroanatomical distribution of NPY receptor subtypes. Using the pharmacological specificity of the NPY receptor subtypes, differential stimulation of GTPgamma[] binding by subtype-specific agonists was used to demonstrate the differential distribution of these subtypes in rat brain. Treatment of rat brain slices with selective agonists for the NPY receptor subtypes in the presence of 2000 microM GDP was used to discriminate populations of NPY receptor subtypes. Activation of a NPY Y1 receptor subtype by human [Leu31Pro34]NPY stimulated GTPgamma[35S] binding in the rank order: frontal cortex>dentate gyrus>inferior colliculus>/=thalamus>hypothalamus. In contrast, NPY Y2/Y5 peptide agonist, human PYY(3-36), stimulated GTPgamma[35S] binding in the rank order: hypothalamus>substantia nigra>hippocampus>frontal cortex>/=inferior colliculus. Stimulation of NPY Y5 receptor subtypes by a NPY Y5 selective agonist, rat/human D-Trp, was shown to stimulate GTPgamma[35S] binding in the hypothalamus and discrete nuclei of the thalamus. Little GTPgamma[35S] binding in the dentate gyrus, frontal cortex, or inferior colliculus was measured following stimulation with D-Trp. Stimulation of GTPgamma[35S] binding by [Leu31Pro34]NPY, but not by the other NPY receptor agonists, was blocked by the selective NPY Y1 receptor antagonist, BIBP 3226. In conclusion, functional coupling at NPY receptor subtypes can be shown in rat brain and populations of NPY receptor subtypes can be anatomically discriminated by NPY agonist stimulation of GTPgamma[35S] binding in rat brain.