Document Detail


Stable expression and pharmacological properties of the human alpha 7 nicotinic acetylcholine receptor.
MedLine Citation:
PMID:  7589218     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The alpha 7 neuronal nicotinic acetylcholine receptor subtype forms a Ca(2+)-permeable homooligomeric ion channel sensitive to alpha-bungarotoxin in Xenopus oocytes. In this study, we have stably and functionally expressed the human alpha 7 cDNA in a mammalian cell line, HEK-293 and examined its pharmacologic properties. [125I] alpha-Bungarotoxin bound to transfected cells with a Kd value of 0.7 nM and a Bmax value of 973 pmoL/mg protein. No specific binding was detected in untransfected cells. Specific binding could be displaced by unlabeled alpha-bungarotoxin (Ki = 0.5 nM) and an excellent correlation was observed between binding affinities of a series of nicotinic cholinergic ligands in transfected cells and those in the human neuroblastoma IMR-32 cell line. Additionally, cell surface expression of alpha 7 receptors was detected by fluorescein isothiocyanate-conjugated alpha-bungarotoxin in transfected cells. Whole cell currents sensitive to blockade by alpha-bungarotoxin, and with fast kinetics of activation and inactivation, were recorded from transfected cells upon rapid application of (-)-nicotine or acetylcholine with EC50 values of 49 microM and 155 microM respectively. We conclude that the human alpha 7 subunit when expressed alone can form functional ion channels and that the stably transfected HEK-293 cell line serves as a unique system for studying human alpha 7 nicotinic receptor function and regulation, and for examining ligand interactions.
Authors:
M Gopalakrishnan; B Buisson; E Touma; T Giordano; J E Campbell; I C Hu; D Donnelly-Roberts; S P Arneric; D Bertrand; J P Sullivan
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  European journal of pharmacology     Volume:  290     ISSN:  0014-2999     ISO Abbreviation:  Eur. J. Pharmacol.     Publication Date:  1995 Aug 
Date Detail:
Created Date:  1995-12-26     Completed Date:  1995-12-26     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  1254354     Medline TA:  Eur J Pharmacol     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  237-46     Citation Subset:  IM    
Affiliation:
Pharmaceutical Products Division, Abbott Laboratories, Abbott Park, IL 60064-3500, USA.
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MeSH Terms
Descriptor/Qualifier:
Acetylcholine / metabolism
Base Sequence
Bungarotoxins / pharmacology
Cell Line
Electrophysiology
Humans
Ion Channel Gating / physiology
Kidney / cytology,  metabolism
Kinetics
Molecular Sequence Data
Nicotine / metabolism
Nicotinic Antagonists / pharmacology
Patch-Clamp Techniques
RNA / metabolism
Receptors, Nicotinic / biosynthesis*,  drug effects*
Recombinant Proteins / biosynthesis,  metabolism
Transfection
Chemical
Reg. No./Substance:
0/Bungarotoxins; 0/Nicotinic Antagonists; 0/Receptors, Nicotinic; 0/Recombinant Proteins; 51-84-3/Acetylcholine; 54-11-5/Nicotine; 63231-63-0/RNA

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