High-Throughput Patch Clamp Screening in Human α6-Containing Nicotinic Acetylcholine Receptors
| Autor: | Abby Sewell, Glenn E. Kirsch, Caiyun Wu, Arianne L. Motter, Fedorov Nikolai, Yuri A. Kuryshev, Lucas C. Armstrong, Michael S. Orr, Zhiqi Liu, Carmine Leggett |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Agonist Patch-Clamp Techniques medicine.drug_class Genetic Vectors Gene Expression Receptors Nicotinic Pharmacology Nucleus accumbens Transfection Biochemistry Cell Line Analytical Chemistry Small Molecule Libraries Nicotine 03 medical and health sciences 0302 clinical medicine Automated patch clamp Drug Discovery medicine Humans Patch clamp nicotinic acetylcholine receptor Cloning Molecular Original Research Acetylcholine receptor Chemistry Electrophysiological Phenomena High-Throughput Screening Assays Protein Subunits Nicotinic acetylcholine receptor 030104 developmental biology Nicotinic agonist ion channel electrophysiological screening Molecular Medicine automated patch clamp Ion Channel Gating 030217 neurology & neurosurgery Biotechnology medicine.drug |
| Zdroj: | Slas Discovery |
| ISSN: | 2472-5552 |
| DOI: | 10.1177/2472555217696794 |
| Popis: | Nicotine, the addictive component of tobacco products, is an agonist at nicotinic acetylcholine receptors (nAChRs) in the brain. The subtypes of nAChR are defined by their α- and β-subunit composition. The α6β2β3 nAChR subtype is expressed in terminals of dopaminergic neurons that project to the nucleus accumbens and striatum and modulate dopamine release in brain regions involved in nicotine addiction. Although subtype-dependent selectivity of nicotine is well documented, subtype-selective profiles of other tobacco product constituents are largely unknown and could be essential for understanding the addiction-related neurological effects of tobacco products. We describe the development and validation of a recombinant cell line expressing human α6/3β2β3V273S nAChR for screening and profiling assays in an automated patch clamp platform (IonWorks Barracuda). The cell line was pharmacologically characterized by subtype-selective and nonselective reference agonists, pore blockers, and competitive antagonists. Agonist and antagonist effects detected by the automated patch clamp approach were comparable to those obtained by conventional electrophysiological assays. A pilot screen of a library of Food and Drug Administration–approved drugs identified compounds, previously not known to modulate nAChRs, which selectively inhibited the α6/3β2β3V273S subtype. These assays provide new tools for screening and subtype-selective profiling of compounds that act at α6β2β3 nicotinic receptors. |
| Databáze: | OpenAIRE |
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