Characterization of Receptors for Cholecystokinin and Related Peptides in Mouse Cerebral Cortex

Abstract: The characteristics of cholecystokinin (CCK) binding to its receptors in a particulate membrane fraction of mouse cerebral cortex were studied by employing biologically active radioiodinated CCK prepared by conjugation with ¹²⁵I-Bolton-Hunter (¹²⁵I-BH) reagent. At 24°C binding was rapid, reversible, and linearly related to protein content. Binding was maximal at acidic pH (6.5) and reduced by the presence of monovalent cations. Under physiological conditions (pH 7.4, 118 mM-NaC1, 4.7 mM-KCl) Scatchard plots of CCK binding were linear with a K _D value of 1.27 nM and binding capacity of 115 fmol/mg protein. Optimal binding required the presence of both Mg²⁺ and EGTA, and was inhibited by the addition of micromolar concentrations of Cu²⁺ (ID₅₀= 30 μM). The cortical receptor recognized all major forms of CCK, with an order of potency of cholecystokinin octapeptide (CCK₈) > CCK > cholecystokinin tetrapeptide (CCK₄). Desulfated cholecystokinin octapeptide (dCCK₈) had a 10-fold lower affhity than CCK₈. Dibutyryl cyclic GMP, a potent competitive inhibitor of CCK binding to receptors in pancreas, was not a specific inhibitor of CCK binding to brain receptors. These present results support the concept that CCK may function as a regulatory peptide in brain, and that the cortical CCK receptor is different from the receptors mediating the peripheral action of CCK.