Demonstration of the dynamic mass redistribution label-free technology as a useful cell-based pharmacological assay for endogenously expressed GABAA receptors

Anders Bue Klein, Mia Nittegaard-Nielsen, Julie T. Christensen, Anas Al-Khawaja, Petrine Wellendorph

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    7 Citations (Scopus)

    Abstract

    Within the continuous quest for the discovery of pharmacologically interesting compounds, the development
    of new and superior drug screening assays is desired. In recent years, the use of label-free techniques
    has paved the way for an alternative high-throughput screening method. An example is the Epic® opticalbased
    biosensor that relies on dynamic mass redistribution (DMR) for detection. So far, DMR assays have
    been mostly used to study G protein-coupled receptor (GPCR) pharmacology. Here, we demonstrate the
    utility of this assay for investigating ligand-gated ion channel receptors. Using the immortalized IMR-32
    neuroblastoma cell line, which expresses relatively high levels of several endogenous GABAA receptor subunits,
    we show that GABA produces concentration-dependent cellular responses that can be measured
    and quantified in real-time. With the aid of the GABAA receptor-specific agonist muscimol and the selective
    antagonists gabazine and bicuculline, we confirm that the data corresponds to that of a GABAA receptor.
    Based on quantitative real-time PCR measurements, the subunits α3, α5, β3 and θ are the most likely candidates
    for integration into functional receptors. Our demonstration that label-free methods such as the Epic
    technology can be used to characterize endogenous GABAA receptors in the IMR-32 cell line is exemplary
    for the superfamily of ligand-gated ion channel receptors, and holds interesting perspectives in relation to
    identifying novel mechanisms of action.
    Original languageEnglish
    JournalMedChemComm
    Volume7
    Pages (from-to)426-432
    Number of pages7
    ISSN2040-2503
    DOIs
    Publication statusPublished - 19 Nov 2015

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