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Fast and Fabulous – Patch Clamp Screening of a7 nAChRs
Author(s) -
Haedo Rodolfo,
Obergrussberger Ali,
Stoelzle Sonja,
Brueggemann Andrea,
Fertig Niels
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.851.4
Subject(s) - homomeric , patch clamp , methyllycaconitine , ion channel , allosteric regulation , neuroscience , pharmacology , chemistry , nicotine , mecamylamine , nicotinic agonist , hek 293 cells , biophysics , nicotinic acetylcholine receptor , protein subunit , electrophysiology , medicine , biology , receptor , biochemistry , gene
To date, nine alpha (a2 to a10) and three Beta subunits (B2 to B4) have been cloned. Given nAChR putative role in cognition and learning & memory, a number of nAChR allosteric modulators (PAMs) have been synthesized and studied as potential therapeutics for conditions such as Alzheimer's disease, schizophrenia and Parkinson's disease. The gold standard for studying ion channels remains the patch clamp technique. One challenge facing the study of homomeric a7 is its extremely fast desensitization. In this study, we have used HEK cells expressing homomeric nAChR a7 subunits or heteromeric a3B4 nAChR and automated patch clamp devices capable of performing whole cell recordings multiple cells simultaneously with high success rates. Characteristic a7 or a3B4 currents were recorded and could be reliably and repetitively activated by addition of nicotine or acetylcholine. Data will be shown for effect of agonists, PAMs and inihibitors, particularly enhancement of a7 currents by PNU‐120596 and inhibition of a3B4 by mecamylamine, a ganglionic blocker with clinically relevant hypotensive actions.