Real-time monitoring of morphological changes in living cells by electronic cell sensor arrays: an approach to study G protein-coupled receptors

Anal Chem. 2006 Jan 1;78(1):35-43. doi: 10.1021/ac051695v.

Abstract

G protein-coupled receptors (GPCRs) constitute important targets for drug discovery against a wide range of ailments including cancer, inflammatory, and cardiovascular diseases. Efforts are underway to screen selective modulators of GPCRs and also to deorphanize GPCRs with unidentified natural ligands. Most GPCR-based cellular screens depend on labeling or recombinant expression of receptor or reporter proteins, which may not capture the true physiology or pharmacology of the GPCRs. In this paper, we describe a noninvasive and label-free assay for GPCRs that can be used with both engineered and nonengineered cell lines. The assay is based on using cell-electrode impedance to measure minute changes in cellular morphology as a result of ligand-dependent GPCR activation. We have used this technology to assay the functional activation of GPCRs coupled to different signaling pathways and have compared it to standard assays. We have used pharmacological modulators of GPCR signaling pathways to demonstrate the specificity of impedance-based measurements. Our data indicate that cell-electrode impedance measurements offer a convenient, sensitive, and quantitative method for assessing GPCR function. Moreover, the noninvasive nature of the readout offers the added advantage of performing multiple treatments in the same well to study events such as desensitization and receptor cross-talk.

MeSH terms

  • Animals
  • Blotting, Western
  • CHO Cells / metabolism
  • CHO Cells / ultrastructure
  • Cricetinae
  • Cricetulus
  • Cyclic AMP / metabolism
  • Dopamine / metabolism
  • Electric Impedance
  • Electronics*
  • HeLa Cells / metabolism
  • HeLa Cells / ultrastructure
  • Histamine / metabolism
  • Humans
  • Immunoprecipitation
  • Inositol Phosphates / metabolism
  • Kinetics
  • Ligands
  • Microscopy, Fluorescence
  • Receptors, Dopamine / genetics
  • Receptors, Dopamine / metabolism
  • Receptors, G-Protein-Coupled / metabolism*
  • Receptors, Histamine / genetics
  • Receptors, Histamine / metabolism
  • Receptors, Vasopressin / genetics
  • Receptors, Vasopressin / metabolism
  • Signal Transduction*
  • Spectrum Analysis
  • Vasopressins / metabolism

Substances

  • Inositol Phosphates
  • Ligands
  • Receptors, Dopamine
  • Receptors, G-Protein-Coupled
  • Receptors, Histamine
  • Receptors, Vasopressin
  • Vasopressins
  • inositol 3-phosphate
  • Histamine
  • Cyclic AMP
  • Dopamine