Characterization of the GABAA receptor in the brain of the adult male bullfrog, Rana catesbeiana

Little is known about the properties of GABA receptors in the amphibian brain. The GABA(A) receptor is widespread in the mammalian brain, and can be specifically labeled with the receptor agonist [H-3]muscimol. The binding of [H-3]muscimol to membrane preparations from the brain of the bullfrog, Rana catesbeiana, was investigated in kinetic, saturation, and inhibition experiments to determine whether this species possessed a GABA(A)-like receptor. Binding of 20 nM [H-3]muscimol to membranes was specific and could be displaced by 1 mM GABA. Association binding curves showed that steady state occurred rapidly, within 2 min, and dissociation occurred within 5 min. The receptor was saturable with a single, high-affinity binding site (K-D = 19.2 nM; B-max = 1.8 pmol/mg protein). Binding of [H-3]muscimol was inhibited in a dose-dependent fashion by muscimol, GABA, bicuculline methiodide, and bicuculline (in order of potency). Baclofen (at doses from 10(-9) to 10(-3) M) failed to displace [H-3]muscimol. The binding characteristics and ligand specificity of [H-3]muscimol binding sites in the bullfrog brain support the hypothesis that this amphibian possesses a GABA(A)-like receptor protein similar to the GABA(A) receptor characterized in mammals. (C) 2003 Elsevier B.V. All rights reserved.