Melatonin receptor-mediated inhibition of cyclic AMP accumulation in chick retinal cell cultures.
Melatonin receptors were characterized in cultured neurons and photoreceptors prepared from chick embryo retina. Cultured cells contained high-affinity 2-[125I]iodomelatonin binding sites (KD = 41.6 pM), similar to those in intact retina. The effects of melatonin and related indoles on cyclic AMP accumulation were examined. Melatonin (10(-7) M) had no effect on basal or K(+)-stimulated cyclic AMP accumulation, but inhibited forskolin-stimulated cyclic AMP accumulation by approximately 50%. Melatonin inhibited forskolin-stimulated cyclic AMP accumulation in the presence or absence of the cyclic nucleotide phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine, suggesting an effect on cyclic AMP synthesis rather than degradation. Half-maximal inhibition was observed at 5.9 x 10(-10) M melatonin. The relative order of potency among melatonin analogues was 2-iodomelatonin > melatonin approximately 6-chloromelatonin > or = 6-hydroxymelatonin > N-acetylserotonin approximately 5-methoxytryptophol > serotonin. The EC50 value for inhibition of cyclic AMP accumulation by 2-iodomelatonin (36.7 pM) was comparable to the KD value for binding of the radioligand, suggesting that the binding sites represent functional receptors. The inhibitory effect of melatonin was antagonized by the putative melatonin antagonists luzindole, N-acetyltryptamine, and N-(2,4-dinitrophenyl)-5-methoxytryptamine, with estimated KB values of 0.12, 0.17, and 1 microM, respectively. At a concentration of 10 microM, N-(2,4-dinitrophenyl)-5-methoxytryptamine significantly inhibited forskolin-stimulated cyclic AMP accumulation when added alone; at 30 microM, luzindole and N-acetyltryptamine also had significant inhibitory effects. The inhibitory effect of melatonin was blocked by pretreatment with pertussis toxin.(ABSTRACT TRUNCATED AT 250 WORDS)
Iuvone
P M
PM
Department of Pharmacology, Emory University School of Medicine, Atlanta, GA 30322.
Gan
J
J
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D008735:5-Methoxytryptamine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000032:analysis / Q000378:metabolism*; D007457:Iodine Radioisotopes; D008550:Melatonin / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology; D009474:Neurons / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D037342:Pertussis Toxin; D010786:Photoreceptor Cells / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D011188:Potassium / Q000494:pharmacology; D011956:Receptors, Cell Surface / Q000378:metabolism / Q000502:physiology*; D044094:Receptors, Melatonin; D012160:Retina / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism*; D015290:Second Messenger Systems / Q000502:physiology; D013329:Structure-Activity Relationship; D014363:Tryptamines / Q000494:pharmacology; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D007457:Iodine Radioisotopes; D011956:Receptors, Cell Surface; D044094:Receptors, Melatonin; D014363:Tryptamines; D010566:Virulence Factors, Bordetella; C053008:ML 23; C057154:luzindole; D005576:Colforsin; D008735:5-Methoxytryptamine; C044556:2-iodomelatonin; D000242:Cyclic AMP; D037342:Pertussis Toxin; D008550:Melatonin; D011188:Potassium; D015056:1-Methyl-3-isobutylxanthine