Stable expression in HEK‐293 cells of the rat α3/β4 subtype of neuronal nicotinic acetylcholine receptor

FEBS Letters

Storch, Alexander²  Kramer, Gert³  Methfessel, Christoph¹  Ebbinghaus, Ullrich²  Maelicke, Alfred²  Stetzer, Eva²  Poteur, Livia²  Schrattenholz, Andre² 

[1] Zentrale Forschung, Bayer AG, 51368 Leverkusen, Germany;Laboratory of Molecular Neurobiology, Institute of Physiological Chemistry and Pathobiochemistry, 6 Duesbergweg, Johannes-Gutenberg University Medical School, 55099 Mainz, Germany;Boehringer-Ingelheim KG, 55216 Ingelheim, Germany

关键词: Stable expression;    HEK cell;    α3/β4 nAChR;    Ganglionic nAChR;    Ca2+ imaging;   

DOI  :  10.1016/S0014-5793(96)01115-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The α3/β4 subtype of neuronal nicotinic acetylcholine receptor (nAChR) was stably expressed in human embryonic kidney (HEK) 293 cells that co-expressed a voltage-gated Ca²⁺ channel. α3/β4-nAChR-expressing clones were identified using the fura-2 Ca²⁺ imaging technique, and were further characterised by single-cell and whole-cell patch-clamp studies. Acetylcholine (ACh) induced fast activating currents which showed desensitisation and inward rectification. The conductance of the ACh-activated channel was 29 pS. The order of potency of the nicotinic agonists tested was . The EC₅₀ value for ACh was 145 μM; the Hill coefficient was close to 2. The currents elicited by ACh were effectively blocked by nicotinic antagonists, but not by the muscarinic antagonist atropine. These properties are comparable to the pharmacological and physiological profile of ganglionic nicotinic receptors and type III currents of cultured hippocampal neurons.

【 授权许可】

Unknown