Optimizing the expression of recombinant αβγ GABAA receptors in HEK293 cells for high-throughput screening

Gilbert, Daniel, Esmaeili, Abolghasem and Lynch, Joseph W. (2009) Optimizing the expression of recombinant αβγ GABAA receptors in HEK293 cells for high-throughput screening. Journal of Biomolecular Screening, 14 1: 86-91. doi:10.1177/1087057108328017


Author Gilbert, Daniel
Esmaeili, Abolghasem
Lynch, Joseph W.
Title Optimizing the expression of recombinant αβγ GABAA receptors in HEK293 cells for high-throughput screening
Formatted title
 Optimizing the expression of recombinant αβγ GABAA receptors in HEK293 cells for high-throughput screening
Journal name Journal of Biomolecular Screening   Check publisher's open access policy
ISSN 1087-0571
1552-454X
Publication date 2009-01-01
Year available 2009
Sub-type Article (original research)
DOI 10.1177/1087057108328017
Open Access Status Not yet assessed
Volume 14
Issue 1
Start page 86
End page 91
Total pages 6
Editor Larry Walker
Place of publication Thousand Oaks, CA , U.S.A.
Publisher Sage Publications
Language eng
Subject C1
110903 Central Nervous System
920111 Nervous System and Disorders
Abstract Despite being important clinical targets, it is not straightforward to reliably express recombinant trimeric αβγ GABA-A receptors (GABAARs) for high-throughput screening. This study therefore sought to devise a simple and reliable means of transiently expressing α1β1γ1 and α1β1γ2 GABAARs in HEK293 cells. Expression efficiencies resulting from 5 different transfection strategies were assessed by flow cytometry and pharmacological analysis using an anion-sensitive yellow fluorescent protein-based assay. PolyFect™ and Effectene™, employed according to the manufacturers' instructions, conferred the strongest and most reliable expression of trimeric αβγ GABAARs. Functional analysis via the yellow fluorescent protein assay revealed dramatic differences in the pharmacological properties of γ1- and γ2-containing receptors, consistent with previous electrophysiological characterizations. The authors conclude that this method of expressing and screening recombinant GABAARs provides an effective means of discovering novel GABAAR modulators for use as therapeutic lead compounds and pharmacological probes.
Keyword Chloride channel
Drug discovery
Yellow fluorescent protein
Yellow fluorescent protein
Inhibitory
Neurotransmission
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: 2010 Higher Education Research Data Collection
Queensland Brain Institute Publications
ERA 2012 Admin Only
 
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Created: Wed, 06 May 2009, 01:56:46 EST by Debra McMurtrie on behalf of Queensland Brain Institute