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Methodological approaches for the study of GABAA receptor pharmacology and functional responses

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Abstract

Inhibitory GABAA receptor ion channels are the target for a wide range of clinically-used therapeutic agents. The complex structural diversity of these ligand-gated channels, revealed by molecular cloning studies, together with increasing requirements for higher-throughput functional assays in drug discovery, has led to the development of a wide range of techniques to examine GABAA receptor pharmacology and function. In the current article we review some of the methodologies which have contributed to the expansion of knowledge in this field. The techniques include: molecular approaches, immunoprecipitation, and immunopurification to study receptor assembly, structure, and functional expression; in situ hybridization, immunocytochemistry, and autoradiography to examine receptor distribution in native tissues; radioligand binding, site-directed mutagenesis, and electrophysiology to examine pharmacology and allosteric modulation; and patch clamp, ion flux, microphysiometry, and a variety of novel fluorescence-based technologies to examine ion-channel function. The use of gene targetting approaches in transgenic mice has also provided important insights into the role of specific GABAA receptor subtypes in vivo. The continuing evolution of novel technologies and assay approaches with appropriate sensitivity and resolution to measure subtle modulation of GABAA ion channels will facilitate ongoing investigation of the physiological functions of these important inhibitory receptors.

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  1. Neuroscience Research Centre, Terlings Park, Merck, Sharp and Dohme Research Laboratories, Eastwick Rd, Harlow, Essex, CM20 2QR, UK

    Alison J. Smith & Peter B. Simpson

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Smith, A.J., Simpson, P.B. Methodological approaches for the study of GABAA receptor pharmacology and functional responses. Anal Bioanal Chem 377, 843–851 (2003). https://doi.org/10.1007/s00216-003-2172-y

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