Stable expression and characterisation of a human ????7 nicotinic subunit chimera: a tool for functional high-throughput screening

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• 作者：Craig ; Peter J. ; Bose ; Suchira ; Zwart ; Ruud ; Beattie ; Ruth E. ; Folly ; Elizabeth A. ; Johnson ; Laura R. ; et. al.
• 关键词：Nicotinic acetylcholine receptor ; ????7 subunit ; ??7/5-HT₃ chimera ; HEK-293 cells ; (Human)
• 刊名：European Journal of Pharmacology
• 出版年：2004
• 出版时间：October 11, 2004
• 年：2004
• 卷：502
• 期：1-2
• 页码：31-40
• 全文大小：487 K

文摘

A chimera comprising the N-terminal region of the human ??7 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 ??7 receptors. A stable HEK-293 cell line expressing the human ??7/mouse 5-HT₃ chimera was established, which showed that the chimera displayed a similar pharmacological profile to wild-type ??7 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 ??7 nicotinic receptors in physiology and disease.