First cloning and functional characterization of a melatonin receptor in fish brain: a novel one?
Melatonin, a neuroendocrine transducer of photoperiod, influences a number of physiological functions and behaviors through specific seven transmembrane domains receptors. We report here the first full-length cloning and functional characterization of a melatonin receptor (P2.6) in a fish, the pike (Teleost). P2.6 encodes a protein that is approximately 80% identical to melatonin receptors previously isolated partially in non-mammals and classified as members of the Mel(1b) subtype; but, it shares only 61% identity with the full-length human Mel(1b) melatonin receptor (hMT2). Expression of P2.6 results in ligand binding characteristics similar to that described for endogenous melatonin receptors. Selective antagonists of the hMT2 (4-phenyl-2-propionamidotetraline and luzindole) were poor competitors of 2-[125I]iodomelatonin binding to the recombinant receptor. In Chinese hamster ovary cells expressing both the cystic fibrosis transmembrane conductance regulator chloride channel and P2.6 receptor, melatonin counteracted the forskolin induced activation of the channel. The results are best explained by a selective inhibition of the adenylyl cyclase. By reverse transcription-polymerase chain reaction, P2.6 mRNA appeared expressed in the optic tectum and, to lesser extent, in the retina and pituitary. In conclusion, these results, together with those of a phylogenetic analysis, suggest that P2.6 might belong to a distinct subtype group within the vertebrate melatonin receptor family.
Gaildrat
Pascaline
P
Département des Neurosciences, CNRS UMR 6558, Université de Poitiers, France.
Becq
Frédéric
F
Falcón
Jack
J
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D015152:Blotting, Northern; D001921:Brain / Q000502:physiology*; D016466:CHO Cells; D003001:Cloning, Molecular; D005576:Colforsin / Q000494:pharmacology; D006224:Cricetinae; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D008550:Melatonin / Q000031:analogs & derivatives* / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D012333:RNA, Messenger / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism*; D044094:Receptors, Melatonin; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013477:Superior Colliculi / Q000502:physiology; D013764:Tetrahydronaphthalenes / Q000378:metabolism
C114459:4-phenyl-2-propionamidotetraline; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D018160:Receptors, Cytoplasmic and Nuclear; D044094:Receptors, Melatonin; D011994:Recombinant Proteins; D013764:Tetrahydronaphthalenes; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005576:Colforsin; C044556:2-iodomelatonin; D008550:Melatonin