[125I]Iodomelatonin-binding sites in the pigeon brain: binding characteristics, regional distribution and diurnal variation.

The binding and pharmacological characteristics of melatonin-binding sites labelled by [125I]iodomelatonin in membrane preparations from the pigeon brain were determined. Specific binding of [125I]iodomelatonin in the membrane preparations of pigeon brain was rapid, stable, saturable and reversible. The [125I]iodomelatonin-binding sites had the following order of pharmacological affinities: melatonin greater than 6-chloromelatonin greater than N-acetylserotonin much greater than 5-hydroxytryptamine greater than tryptamine greater than 5-methoxytryptophol, greater than 1-acetylindole-3-carboxytryptamine, 5-hydroxyindole-3-acetic acid, L-tryptophan and 3-acetylindole. Compounds known to act on serotonin receptors, adrenoceptors or cholinoceptors were inactive compared with melatonin. Of the various brain regions studied, melatonin binding was greatest in the hypothalamus, intermediate in the mid-brain, pons-medulla and telencephalon, and low in the cerebellum. Subcellular fraction studies indicated that 39% of the binding was located in the mitochondrial fraction, 34% in the nuclear fraction, 21% in the microsomal fraction and 5.6% in the cytosol fraction. Scatchard analysis of the membrane preparations revealed a dissociation constant (Kd) of 206.3 +/- 57.9 pmol/l and a total number of binding sites (Bmax) of 26.7 +/- 1.9 fmol/mg protein in the middle of the light period (mid-light). In addition, saturation studies demonstrated that [125I]iodomelatonin-binding sites in pigeon brain membrane preparations were 36.2% higher at mid-light (26.7 +/- 1.9 fmol/mg protein) than in the middle of the dark period (19.6 +/- 1.25 fmol/mg protein), with no significant variation in their binding affinities.