SELECTIVE IN-VIVO LABELING OF BRAIN 5-HT1A-RECEPTORS BY [H-3] WAY-100635 IN THE MOUSE

The novel selective 5-HT1A receptor antagonist radioligand [H-3]WAY 100635 ([O-methy-H-3]N-(2-(4-(2-methoxyphenyl)-1-piperazinyl)ethyl)-N-(2-pyridyl)cyclohexane oxamide) was injected i.v. to mice in an attempt to label in vivo central 5-HT1A receptors. Although 5 min after the i.v, injection of [H-3]WAY 100635 (4-7.6 mu Ci per mouse) the amount of tritium found in the whole brain only accounted for 1.5-1.8% of the injected radioactivity, regional differences in H-3 accumulation already corresponded to those of 5-HT1A receptor density. Optimal data were obtained 1 h after [H-3]WAY 100635 injection as the distribution of H-3 in brain was exactly that of 5-HT1A receptor binding sites in mouse brain sections labelled in vitro with [H-3]WAY 100635. In particular, high level of labelling was found in the lateral septum, gyrus dentatus and CA1 area of Ammon's horn in the hippocampus, dorsal raphe nucleus and entorhinal cortex. No labelling was found in the substantia nigra, and H-3 accumulated in the cerebellum represented only 12-14% of that found in the hippocampus. Pretreatment with various drugs indicated that only 5-HT1A receptor ligands were able to decrease the accumulation of H-3 in all the brain areas examined except in the cerebellum. Assuming that only non-specific binding took place in the latter structure, it was possible to calculate the ID50 values of 5-HT1A receptor agonists (8-OH-DPAT (8-hydroxy-2-(di-n-propylamino)tetralin) S 14506 (1-[2-(4-fluorobenzoylamino)ethyl]-4-(7-methoxynaphthyl)piperazine) and S 20499 ((+)-4-[N-(5-methoxy-chroman-3-yl)-N-propylamino]butyl-8-azaspiro -(4,5)-decane-7,9-dione)) and antagonists (spiperone (-)-tertatolol, (+)-WAY 100135 (N-tert-butyl-3,4-(2-methoxy-phenyl)piperazin-1-yl-2-phenyl-propanamide)) as inhibitors of H-3 accumulation in the hippocampus of [H-3]WAY 100635-injected mice. Comparison of these values with the in vitro affinity of the same ligands for hippocampal 5-HT1A receptors revealed marked variations in the capacity of 5-HT1A receptor agonists and antagonists to reach the brain when injected via the subcutaneous route in mice.