TY - JOUR

T1 - Demonstration of the dynamic mass redistribution label-free technology as a useful cell-based pharmacological assay for endogenously expressed GABAA receptors

AU - Klein, Anders Bue

AU - Nittegaard-Nielsen, Mia

AU - Christensen, Julie T.

AU - Al-Khawaja, Anas

AU - Wellendorph, Petrine

PY - 2015/11/19

Y1 - 2015/11/19

N2 - Within the continuous quest for the discovery of pharmacologically interesting compounds, the developmentof new and superior drug screening assays is desired. In recent years, the use of label-free techniqueshas paved the way for an alternative high-throughput screening method. An example is the Epic® opticalbasedbiosensor that relies on dynamic mass redistribution (DMR) for detection. So far, DMR assays havebeen mostly used to study G protein-coupled receptor (GPCR) pharmacology. Here, we demonstrate theutility of this assay for investigating ligand-gated ion channel receptors. Using the immortalized IMR-32neuroblastoma 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 measuredand quantified in real-time. With the aid of the GABAA receptor-specific agonist muscimol and the selectiveantagonists 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 candidatesfor integration into functional receptors. Our demonstration that label-free methods such as the Epictechnology can be used to characterize endogenous GABAA receptors in the IMR-32 cell line is exemplaryfor the superfamily of ligand-gated ion channel receptors, and holds interesting perspectives in relation toidentifying novel mechanisms of action.

AB - Within the continuous quest for the discovery of pharmacologically interesting compounds, the developmentof new and superior drug screening assays is desired. In recent years, the use of label-free techniqueshas paved the way for an alternative high-throughput screening method. An example is the Epic® opticalbasedbiosensor that relies on dynamic mass redistribution (DMR) for detection. So far, DMR assays havebeen mostly used to study G protein-coupled receptor (GPCR) pharmacology. Here, we demonstrate theutility of this assay for investigating ligand-gated ion channel receptors. Using the immortalized IMR-32neuroblastoma 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 measuredand quantified in real-time. With the aid of the GABAA receptor-specific agonist muscimol and the selectiveantagonists 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 candidatesfor integration into functional receptors. Our demonstration that label-free methods such as the Epictechnology can be used to characterize endogenous GABAA receptors in the IMR-32 cell line is exemplaryfor the superfamily of ligand-gated ion channel receptors, and holds interesting perspectives in relation toidentifying novel mechanisms of action.

U2 - 10.1039/c5md00442j

DO - 10.1039/c5md00442j

M3 - Journal article

VL - 7

SP - 426

EP - 432

JO - MedChemComm

JF - MedChemComm

SN - 2040-2503

ER -