This Article Full Text (PDF) References Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Gilbert, D. Articles by Lynch, J. W. Search for Related Content PubMed PubMed Citation Articles by Gilbert, D. Articles by Lynch, J. W. Social Bookmarking                         What's this?

Optimizing the Expression of Recombinant β GABA_A Receptors in HEK293 Cells for High-Throughput Screening

Queensland Brain Institute, University of Queensland, Brisbane, Australia, School of Biomedical Sciences, University of Queensland, Brisbane, Australia

Abolghasem Esmaeili

Queensland Brain Institute, University of Queensland, Brisbane, Australia, Department of Biology, University of Isfahan, Isfahan, Iran

Joseph W. Lynch

Queensland Brain Institute, University of Queensland, Brisbane, Australia, School of Biomedical Sciences, University of Queensland, Brisbane, Australia, j.lynch{at}uq.edu.au

Despite being important clinical targets, it is not straightforward to reliably express recombinant trimeric β GABA-A receptors (GABA_ARs) for high-throughput screening. This study therefore sought to devise a simple and reliable means of transiently expressing 1β11 and 1β12 GABA_ARs 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^TM and Effectene^TM, employed according to the manufacturers' instructions, conferred the strongest and most reliable expression of trimeric β GABA_ARs. Functional analysis via the yellow fluorescent protein assay revealed dramatic differences in the pharmacological properties of 1- and 2-containing receptors, consistent with previous electrophysiological characterizations. The authors conclude that this method of expressing and screening recombinant GABA_ARs provides an effective means of discovering novel GABA_AR modulators for use as therapeutic lead compounds and pharmacological probes. (Journal of Biomolecular Screening 2009:86-91)

Key Words: chloride channel • drug discovery • yellow fluorescent protein • inhibitory • neurotransmission

Journal of Biomolecular Screening, Vol. 14, No. 1, 86-91 (2009)
DOI: 10.1177/1087057108328017


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				D. F. Gilbert, T. Meinhof, R. Pepperkok, and H. Runz DetecTiff(C): A Novel Image Analysis Routine for High-Content Screening Microscopy J Biomol Screen, September 1, 2009; 14(8): 944 - 955. [Abstract] [PDF]