Pharmacological characterization of melatonin binding sites in Syrian hamster hypothalamus

The radioligand [125I]iodomelatonin was used to study melatonin binding sites in Syrian hamster hypothalamus and hippocampus. Scatchard analysis revealed a single binding site with nanomolar affinity in hypothalamus (Kd = 1.8 +/- 0.3 nM, Bmax = 75 +/- 7 fmol/mg protein; n = 4) and hippocampus (Kd = 2.2 +/- 0.2 nM, Bmax = 49 +/- 5 fmol/mg protein; n = 4). The Kd value calculated from the association and dissociation rate constants in hypothalamus was (k-1/k1) = 2.4 nM. Regional studies revealed that the highest binding of [125I]iodomelatonin occurs in the hypothalamus. Only indoles structurally related to melatonin exhibited significant affinity at this site. Prazosin was found to be a potent inhibitor of [125I]iodomelatonin binding in all brain regions studied. The pharmacological profile of this binding site indicated it to be unique, since serotonergic, dopaminergic and adrenergic drugs (other than prazosin) did not have appreciable affinity for it. Although saturation studies revealed only one binding site, the low Hill coefficients obtained for several inhibitors suggest that [125I]iodomelatonin labels multiple sites (or affinity states) in the hypothalamus.