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Journal of Neuroscience, Vol 9, 2581-2590, Copyright © 1989 by Society for Neuroscience
Localization and characterization of melatonin receptors in rodent brain by in vitro autoradiography
DR Weaver, SA Rivkees and SM Reppert
Laboratory of Developmental Chronobiology, Massachusetts General Hospital, Boston.
Little is known of the neural sites of action for the pineal hormone,
melatonin. Thus, we developed an in vitro autoradiographic method using
125I-labeled melatonin (I-MEL) to study putative melatonin receptors in
rodent brain. We first determined optimal in vitro labeling conditions for
autoradiographic detection of I-MEL binding sites in rat median eminence,
the most intensely labeled area in the rat brain. We then assessed the
pharmacologic and kinetic properties of I-MEL binding sites in rat median
eminence by quantitative autoradiography. These sites have high affinity
for I-MEL (equilibrium dissociation constant = 43 pM). I-MEL binding was
inhibited by nanomolar concentrations of melatonin or 6-chloromelatonin,
but was not inhibited by serotonin, dopamine, or norepinephrine (100
microM). These results suggest that I- MEL binding sites identified by in
vitro autoradiography represent specific, high-affinity melatonin
receptors. Studies of the distribution of I-MEL binding in rat, Syrian
hamster, and Djungarian hamster brain confirm that the median eminence and
suprachiasmatic nucleus are major sites of I-MEL binding in rodent brain;
other brain areas labeled in one or more of these species were the thalamus
(paraventricular, anteroventral, and reuniens nuclei, nucleus of the stria
medullaris, and medial part of the lateral habenular nucleus), hypothalamus
(dorsomedial nucleus), subiculum, and area postrema. The presence of
putative melatonin receptors in the suprachiasmatic nuclei and median
eminence of these rodent species suggests that these brain regions are
important loci for melatonin effects on circadian rhythms and reproduction.
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