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Stable expression and characterisation of a human alpha 7 nicotinic subunit chimera: a tool for functional high-throughput screening
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Dr Emma Bell
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
European Journal of Pharmacology
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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 Xenopus 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 pharmacological 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 the role of alpha7 nicotinic receptors in physiology and disease.
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