Neuropeptide Y (NPY) binding sites in rat brain labeled with 125I-Bolton-Hunter NPY: Comparative potencies of various polypeptides on brain NPY binding and biological responses in the rat vas deferens

Abstract The binding of biologically active 125 I-Bolton-Hunter (BH)-NPY to rat brain membranes was saturable and reversible and regulated by inorganic cations and guanyl nucleotides consistent with other neurotransmitter receptor systems. The concentration of specific 125 I-NPY binding differed in various brain regions, being highest in the hippocampus and lowest in the cerebellum. Scatchard analysis of 125 I-NPY binding showed a single class of receptor sites with a K d = 0.1 nM and B max of 3 pmole/g tissue in hippocampus. Peptide YY, porcine and human NPY inhibited the specific 125 I-BH-NPY binding with IC 50 values of 50–120 pM. In contrast, human NPY free acid and pancreatic polypeptides from human (HPP), rat (RPP) and avian (APP) sources were much weaker (IC 50 ≥ 300 nM). The rank order of potencies for NPY analogs and the inactivity of APP and HPP fragment (31–36) on brain binding appeared to correlate with their relative activities in inhibiting contractions of the field-stimulated rat vas deferens. However, PYY, HPP and RPP exhibited activity in the field-stimulated rat vas deferens indicative of a possible action upon sites distinct from the brain NPY binding site.