Stable expression and characterisation of a human α7 nicotinic subunit chimera:: a tool for functional high-throughput screening

被引:22
作者
Craig, PJ
Bose, S
Zwart, R
Beattie, RE
Folly, EA
Johnson, LR
Bell, E
Evans, NM
Benedetti, G
Pearson, KH
McPhie, GI
Volsen, SG
Millar, NS
Sher, E
Broad, LM
机构
[1] Eli Lilly & Co, Lilly Res Ctr Ltd, Windlesham GU20 6PH, Surrey, England
[2] UCL, Dept Pharmacol, London WC1E 6BT, England
关键词
nicotinic acetylcholine receptor; alpha; 7; subunit; 7/5-HT3; chimera; HEK-293; cells; (human);
D O I
10.1016/j.ejphar.2004.08.042
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
A chimera comprising the N-terminal region of the human alpha7 nicotinic acetylcholine receptor, fused to the transmembrane/C-terminal domains of the mouse serotonin 5-HT3 receptor, was constructed. Injection of the chimera cDNA into Xellopus oocytes, or transient transfection in human embryonic kidney (HEK-293) cells, resulted in the expression of functional channels that were sensitive to nicotinic acetylcholine, but not serotonin receptor ligands. In both systems, the responses obtained from chimeric receptors inactivated more slowly than those recorded following activation of wild-type alpha7 receptors. A stable HEK-293 cell line expressing the human alpha7/mouse 5-HT3 chimera was established, which showed that the chimera displayed a similar plianiiaeological profile to wild-type alpha7 receptors. Use of this chimera in high-throughput screening may enable the identification of novel pharmacological agents that will help to define further :he role of alpha7 nicotinic receptors in physiology and disease. (C) 2004 Elsevier B.V. All rights reserved.
引用
收藏
页码:31 / 40
页数:10
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