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 an d presumably exerts its activities through a series of receptors among whic h 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 t hat 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, m ost importantly, that it was measurable at 20 degrees while it has been ini tially described and thoroughly studied using 2-[I-125]-iodomelatonin at 4 degrees. Under these novel conditions, binding to MT3 could be traced witho ut cross-reactivity with MT1 and MT2 receptors and, moreover, under conditi ons similar to those used to measure MT3/QR(2) catalytic activity. The phar macology established here on hamster kidney samples using the reference com pounds remained essentially as already described using other experimental c onditions. A new series of compounds with nanomolar affinity for the MT3 bi nding site and a high MT3 selectivity versus MT1 and MT2 is reported. In ad dition, we further document the MT3/QR(2) binding site by demonstrating tha t it was widely distributed among mammals, although inter-species and inter -tissues differences exist. The present report details new experimental con ditions for the pharmacological study of melatonin-sensitive QR(2) isoforms , and suggests that, in addition to an already demonstrated inter-species d ifference, inter-tissues differences in QR(2) sensitivity to melatonin may exist in primates and, therefore, represent an original and interesting rou te of investigation on the effect of melatonin on MT3/QR(2). (C) 2001 Elsev ier Science Inc. All rights reserved.