Serotonin antagonizes the human neuronal alpha7 nicotinic acetylcholine receptor and becomes an agonist after L248T alpha7 mutation.
The effects of serotonin (5-hydroxytryptamine or 5HT) on chick alpha7 nicotinic receptors have already been described. However similar studies on human alpha7 receptors have been lacking. To begin to fill this deficiency, studies were made on wild-type and mutant human alpha7 (halpha7) receptors expressed in Xenopus oocytes or human BOSC 23 cells. In oocytes wild-type halpha7 receptors were blocked by 5HT, and this block was voltage-dependent. In contrast, 5HT acted as an agonist on halpha7-mutant receptors (L248T). Outside-out membrane-patches from BOSC 23 cells expressing halpha7-mutant receptors exhibited spontaneous channel openings of two conductance levels (59 pS and 76 pS) and short mean open time (0.9 ms). halpha7-Mutant channels activated by nicotine or 5HT displayed similar conductances and high Ca(2+) permeability; but longer duration (2.7 ms) than the spontaneous openings. Mutations at Cys190 and Cys191, in the extracellular N-terminus of the human alpha7 gene, did not prevent receptor expression and incorporation in the oocyte membrane (determined by alpha-bungarotoxin binding). However, both 5HT and nicotine were incapable of gating the channels, indicating that the mutated Cys residues are in, or near, the 5HT- and nicotine-binding site. This is the first report that alpha7 receptors have spontaneous openings; and that 5HT is an agonist of halpha7-mutant receptors, and an antagonist of halpha7-wild-type receptors, through interactions at, or near the acetylcholine-binding sites.
Fucile
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Istituto Pasteur-Fondazione Cenci Bolognetti, Dipartimento di Fisiologia Umana e Farmacologia, Universitá di Roma La Sapienza, Viale Regina Elena 324, I-00161 Rome, Italy.
Palma
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Eusebi
F
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Miledi
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R
D000109:Acetylcholine / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites / Q000187:drug effects / Q000235:genetics; D001921:Brain / Q000378:metabolism*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions / Q000502:physiology; D005260:Female; D006801:Humans; D007473:Ion Channels / Q000187:drug effects / Q000235:genetics; D008564:Membrane Potentials / Q000187:drug effects / Q000235:genetics; D009154:Mutation / Q000187:drug effects* / Q000235:genetics; D009474:Neurons / Q000378:metabolism*; D009538:Nicotine / Q000494:pharmacology; D018733:Nicotinic Antagonists / Q000494:pharmacology; D009865:Oocytes; D011978:Receptors, Nicotinic / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D012701:Serotonin / Q000378:metabolism / Q000494:pharmacology*; D013570:Synaptic Membranes / Q000187:drug effects* / Q000378:metabolism; D009435:Synaptic Transmission / Q000187:drug effects* / Q000502:physiology; D014982:Xenopus laevis; D064569:alpha7 Nicotinic Acetylcholine Receptor
C579167:Chrna7 protein, human; D007473:Ion Channels; D018733:Nicotinic Antagonists; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor; D012701:Serotonin; D009538:Nicotine; D000109:Acetylcholine