Cloning, expression, and characterization of ferret 5-HT(3) receptor subunit

Abstract

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.