2-(2-benzofuranyl)-2-imidazoline and cirazoline

The novel selective imidazoline radioligand [3H]2-(2-benzofuranyl)-2-imidazoline (2-BFI) was used to characterize and assess further the nature of I2-imidazoline receptors in rat brain and liver. In the cerebral cortex, 2-BFI displayed high affinity (Ki = 9.8 nM) for a single class of [3H]2-BFI binding sites. Other imidazoline/guanidine compounds (e.g. aganodine, cirazoline and idazoxan) displayed biphasic competition curves, indicating the existence of high (KiH = 2.9-78 nM; R(H) = 61-83%) and low (KiL = 4.7-158 microM) affinity sites. The pharmacological profile for [3H]2-BFI binding (aganodine > cirazoline > 2-BFI >> clonidine > amiloride >> efaroxan) was typical of that for I2-sites. This profile was almost identical to that obtained against [3H]idazoxan (correlation between pKi values, r = 0.97) which indicated that the sites characterized with [3H]2-BFI in brain corresponded to I2-imidazoline receptors. The low affinity of amiloride against [3H]2-BFI (Ki = 900 nM) further indicated that these brain I2-sites belong to the I2B-subtype. [3H]2-BFI binding sites (Bmax = 72 fmol/mg protein) in brain were differentially modulated by treatment (7 days) with cirazoline (up-regulation: 25%) and the MAO inhibitor phenelzine (down-regulation: 31%), indicating that these I2-sites are regulated in vivo, as is the case for those labelled by [3H]idazoxan. Chronic treatment with 2-phenylethylamine, a phenelzine metabolite and endogenous amine, did not alter the density of brain of I2-imidazoline receptors labelled by [3H]idazoxan. Preincubation of liver membranes with the MAO inhibitor clorgyline (10(-7) M) abolished the binding of [3H]Ro 41-1049 (N-(2-aminoethyl)-5-(m-fluorophenyl)-4-thiazole carboxamide) to MAO-A, but it did not alter the binding of [3H]Ro 19-6327 (N-(2-aminoethyl)-5-chloro-2-pyridine carboxamide) to MAO-B or that of [3H]2-BFI to I2-sites. At 10(-4) M it also abolished MAO-B sites, but a substantial proportion of I2-sites (40%) remained intact. Preincubation of liver membranes at 60 degrees C also abolished MAO-A/B sites, whereas still 22% of I2-sites remained. The results indicate that [3H]2-BFI is a good tool for the identification of I2-imidazoline receptors and suggest further that certain I2-sites and MAO are different proteins.

Topics: Adrenergic beta-Antagonists; Affinity Labels; Animals; Benzofurans; Binding, Competitive; Brain; Cerebral Cortex; Idazoxan; Imidazoles; Imidazoline Receptors; In Vitro Techniques; Isoenzymes; Ligands; Liver; Male; Monoamine Oxidase; Monoamine Oxidase Inhibitors; Phenelzine; Protein Binding; Rats; Rats, Sprague-Dawley; Receptors, Drug