Gamma-aminobutyric acid (GABA) is the main inhibitory neurotransmitter in the vertebrate brain. GABA activates both ionotropic (GABA A ) and metabotropic (GABA B ) receptors in mammals. Whether non-mammalian vertebrates possess receptors with similar characteristics is not well understood. We used a mammalian GABA B -specific antagonist to determine the pharmacology of putative receptors in the brain of an anuran amphibian, the male bullfrog (Rana catesbeiana). Receptor binding assays with the antagonist [ 3 H]CGP54626 revealed a single class of high affinity binding sites (with a K D of 2.97 nM and a B max of 2619 fmol/mg protein). Binding was time- and temperature-dependent, saturable and specific. Specific binding of [ 3 H]CGP54626 was inhibited by several mammalian GABA B receptor agonists and antagonists. The rank order potency of agonists was: GABA = SKF97541 > (R)-Baclofen > 3-APPA. The rank order for antagonists was: CGP54626 = CGP55845 > CGP52432 > CGP35348. The GABA A receptor ligands muscimol and SR95531 had very low affinity for [ 3 H]CGP54626 binding sites, while bicuculline compounds had no affinity. Binding of GABA was positively modulated by CGP7930. Taurine did not allosterically modulate GABA binding but did inhibit [ 3 H]CGP54626 binding in a linear fashion. Bullfrog brain thus possesses binding sites with significant similarity to mammalian GABA B receptors. These receptors differ from mammalian receptors, however, in dissociation kinetics, ligand specificity and allosteric modulation.