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- W2000747496 abstract "Center for Proteomic Chemistry, Novartis Institutes for BioMedical Research Inc., 250 Massachusetts Avenue Cambridge, MA 02139, USA.QPatch HTX is an automated patch clamp system with greatly increased throughput over conventional patch clamp methods. In this study, we evaluated the suitability of this system for the characterization of the fast desensitizing ligand-gated ion channel coupled with alpha1 nicotinic acetylcholine receptors (α1nAChR). HEK293 cells stably expressing human α1nAChR were obtained from Millipore. All experiments were carried out with the QPatch HTX, which performs 48 parallel and independent patch-clamp recordings on a disposable QPlate X with 10 holes per measurement site. Brief applications of acetylcholine (ACh) produced fast inward currents characteristic of the α1nAChR recorded by conventional patch clamp in heterologous expression systems and the native channel. The EC50 values for ACh ranged from 3 to 10 µM (n=10). Nicotine and epibatidine also elicited inward currents with EC50 values of 110 µM and 4 µM, respectively (n=6 & 12). The ACh-induced current was resistant to bath applications of atropine up to 1 µM and muscarine did not elicit a response, showing that the ACh-induced inward current is mediated by nAChR. Non expressing HEK293 cells did not respond to ACh. The ACh-mediated currents were blocked by α-bungarotoxin and pancuronium in a dose dependent manner (IC50= 80 and 0.8 nM, respectively).In conclusion, this study demonstrated that fast desensitizing ligand-gated ion channel can be efficiently targeted with QPatch HTX system for characterization of physiological and pharmacological properties." @default.
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- W2000747496 date "2011-02-01" @default.
- W2000747496 modified "2023-09-26" @default.
- W2000747496 title "Assay Development and Validation for the Muscle Nicotinic Receptor Using the Qpatch HTX™ Automated Patch Clamp System" @default.
- W2000747496 doi "https://doi.org/10.1016/j.bpj.2010.12.1710" @default.
- W2000747496 hasPublicationYear "2011" @default.
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