Comparative pharmacological studies of melatonin receptors:: MT1, MT2 and MT3/QR2.: Tissue distribution of MT3/QR2

The neurohormone melatonin is the central switch of the circadian rhythm and presumably exerts its activities through a series of receptors among which MT1 and MT2 have been widely studied. The third binding site of melatonin, MT3, has been recently characterized as a melatonin-sensitive Term of the quinone reductase 2 (QR(2), EC 1.6.99.2). In the present work, we showed that the binding of melatonin at MT3/QR(2)was better described with 2-[I-125]-iodomethoxy-carbonylamino-N-acetyltryptamine (2-[I-125]-I-MCA-NAT) and, most importantly, that it was measurable at 20 degrees while it has been initially described and thoroughly studied using 2-[I-125]-iodomelatonin at 4 degrees. Under these novel conditions, binding to MT3 could be traced without cross-reactivity with MT1 and MT2 receptors and, moreover, under conditions similar to those used to measure MT3/QR(2) catalytic activity. The pharmacology established here on hamster kidney samples using the reference compounds remained essentially as already described using other experimental conditions. A new series of compounds with nanomolar affinity for the MT3 binding site and a high MT3 selectivity versus MT1 and MT2 is reported. In addition, we further document the MT3/QR(2) binding site by demonstrating that it was widely distributed among mammals, although inter-species and inter-tissues differences exist. The present report details new experimental conditions for the pharmacological study of melatonin-sensitive QR(2) isoforms, and suggests that, in addition to an already demonstrated inter-species difference, inter-tissues differences in QR(2) sensitivity to melatonin may exist in primates and, therefore, represent an original and interesting route of investigation on the effect of melatonin on MT3/QR(2). (C) 2001 Elsevier Science Inc. All rights reserved.