Cloning, expression, and characterization of ferret 5-HT(3) receptor subunit.
Ferrets (Mustela putorius furo) are useful animals for determining anti-emetic activity via 5-HT(3) receptors in vivo. We isolated a cDNA encoding the 5-hydroxytryptamine (5-HT) type 3A receptor subunit (5-HT(3A)) from ferret colon, expressed it in a human embryonic kidney cell line and determined its pharmacological properties. The open reading frame of the isolated cDNA encoded a 483-amino acid protein, corresponding to the shorter splice variant of 5-HT(3A) receptors. Splice variants were no longer detected by reverse transcriptase-polymerase chain reaction. The ferret 5-HT(3A) receptor exhibits a high degree of amino acid sequence identity (>/=80%) to that of other species. Binding studies demonstrated the following rank order of potency for agonists: meta-chlorophenylbiguanide (mCPBG)>2-methyl-5-hydroxytryptamine (2-Me-5-HT)=5-HT, and for antagonists: ondansetron=tropisetron>(+)-tubocurarine>metoclopramide. Electrophysiological studies revealed that mCPBG was a partial agonist and 2-Me-5-HT was an almost fully effective agonist compared to 5-HT.
Mochizuki
S
S
Molecular Medicine Laboratories, Institute for Drug Discovery Research, Yamanouchi Pharmaceutical Co., Ltd., 21 Miyukigaoka, Tsukuba, 305-8585, Ibaraki, Japan. mochizuk@yamanouchi.co.jp
Watanabe
T
T
Miyake
A
A
Saito
M
M
Furuichi
K
K
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001645:Biguanides / Q000494:pharmacology; D001667:Binding, Competitive; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D005289:Ferrets; D015870:Gene Expression; D006801:Humans; D007093:Imidazoles / Q000378:metabolism; D007211:Indoles / Q000378:metabolism; D000075723:Kruppel-Like Factor 6; D051741:Kruppel-Like Transcription Factors; D008564:Membrane Potentials / Q000187:drug effects; D008969:Molecular Sequence Data; D017294:Ondansetron / Q000494:pharmacology; D010879:Piperazines; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011869:Radioligand Assay; D051381:Rats; D011985:Receptors, Serotonin / Q000235:genetics*; D044406:Receptors, Serotonin, 5-HT3; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012701:Serotonin / Q000031:analogs & derivatives* / Q000494:pharmacology; D012702:Serotonin Antagonists / Q000494:pharmacology; D017366:Serotonin Receptor Agonists / Q000494:pharmacology; D014018:Tissue Distribution; D015534:Trans-Activators; D014316:Tritium; D014403:Tubocurarine / Q000494:pharmacology
D001645:Biguanides; D018076:DNA, Complementary; D007093:Imidazoles; D007211:Indoles; C105465:KLF6 protein, human; C491357:Klf6 protein, rat; D000075723:Kruppel-Like Factor 6; D051741:Kruppel-Like Transcription Factors; D010879:Piperazines; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D011985:Receptors, Serotonin; D044406:Receptors, Serotonin, 5-HT3; D012702:Serotonin Antagonists; D017366:Serotonin Receptor Agonists; D015534:Trans-Activators; D014316:Tritium; C059068:GR 65630; D012701:Serotonin; D017294:Ondansetron; C045918:2-methyl-5-HT; C065734:1-(3-chlorophenyl)biguanide; C015068:1-(3-chlorophenyl)piperazine; D014403:Tubocurarine