title: 2-[125I]iodomelatonin labels sites with identical pharmacological characteristics in chicken brain and chicken retina.
abstract: The binding and pharmacological characteristics of the melatonin site labeled by the radioligand 2-[125I]iodomelatonin in chicken brain membranes were determined and compared with those of the melatonin site of chicken retinal membranes. The specific binding of 2-[125I]iodomelatonin to chicken brain membranes was found to be stable, saturable, reversible and of high affinity. Scatchard analysis of the binding revealed an affinity constant (Kd) of 344 +/- 24 pM (n = 4) and a total number of binding sites (Bmax) of 57.6 +/- 10.1 fmol/mg protein (n = 4). The Kd value correspond closely with that found in kinetic studies (Kd = 407 pM) and that reported in chicken retinal membranes. Competition experiments were carried out with various compounds revealing the following order of pharmacological affinities: 6-chloromelatonin greater than or equal to 2-iodomelatonin greater than melatonin greater than 2-methyl-6,7-dichloromelatonin greater than 6-hydroxymelatonin greater than N-acetyl-5-hydroxytryptamine greater than luzindole greater than N-acetyl-5-methoxykynurenamine greater than 6-methoxymelatonin greater than N-acetyltryptamine greater than 5-methoxytryptamine greater than 5-hydroxytryptamine greater than 5-methoxy-N,N-dimethyltryptamine greater than 5-methoxytryptophol. This order of pharmacological affinities is identical to that found in chicken retinal membranes. Correlation between affinity constants for various melatonin receptor agonists and putative melatonin receptor antagonists obtained in chicken brain and retinal membranes yielded a correlation coefficient (r) of 0.966 (slope = 0.652, n = 14, P less than 0.01). We conclude that the high affinity site labeled by 2-[125I]iodomelatonin in chicken brain membranes has identical binding and pharmacological characteristics to the ML-1 melatonin receptor site previously described in chicken retinal membranes.
authors.affiliation: Department of Pharmacology, Northwestern University Medical School, Chicago, IL 60611.
authors.forename: M L
authors.initials: ML
authors.lastname: Dubocovich
authors.forename: G
authors.initials: G
authors.lastname: Shankar
authors.forename: M
authors.initials: M
authors.lastname: Mickel
mesh_terms: D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects; D001923:Brain Chemistry / Q000187:drug effects*; D002645:Chickens; D006224:Cricetinae; D066298:In Vitro Techniques; D007211:Indoles / Q000378:metabolism; D007457:Iodine Radioisotopes; D008550:Melatonin / Q000031:analogs & derivatives* / Q000378:metabolism / Q000493:pharmacokinetics; D008566:Membranes / Q000187:drug effects / Q000378:metabolism; D044094:Receptors, Melatonin; D017981:Receptors, Neurotransmitter / Q000378:metabolism*; D012160:Retina / Q000187:drug effects / Q000378:metabolism*
chemical_list: D007211:Indoles; D007457:Iodine Radioisotopes; D044094:Receptors, Melatonin; D017981:Receptors, Neurotransmitter; C044556:2-iodomelatonin; D008550:Melatonin