A FLIPR assay for evaluating agonists and antagonists of GPCR heterodimers

Methods Mol Biol. 2013:995:43-54. doi: 10.1007/978-1-62703-345-9_4.

Abstract

Calcium signaling plays a major role in the function of cells. Measurement of intracellular calcium mobilization is a robust assay that can be performed in a high-throughput manner to study the effect of compounds on potential drug targets. Pharmaceutical companies frequently use calcium signaling assays to screen compound libraries on G-protein-coupled receptors (GPCRs). In this chapter we describe the application of FLIPR technology to the evaluation of GPCR-induced calcium mobilization. We also include the implications of GPCR hetero-oligomerization and the identification of heteromeric receptors as novel drug targets on high-throughput calcium screening.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Benzylidene Compounds / pharmacology*
  • Calcium Signaling
  • Cell Culture Techniques
  • Drug Evaluation, Preclinical / methods*
  • Enkephalin, D-Penicillamine (2,5)- / pharmacology*
  • HEK293 Cells
  • High-Throughput Screening Assays / methods*
  • Humans
  • Naltrexone / analogs & derivatives*
  • Naltrexone / pharmacology
  • Receptors, G-Protein-Coupled / agonists
  • Receptors, G-Protein-Coupled / antagonists & inhibitors
  • Receptors, G-Protein-Coupled / biosynthesis
  • Receptors, Opioid, delta / agonists*
  • Receptors, Opioid, delta / antagonists & inhibitors
  • Receptors, Opioid, delta / biosynthesis
  • Recombinant Proteins / agonists
  • Recombinant Proteins / antagonists & inhibitors
  • Recombinant Proteins / biosynthesis
  • Spectrometry, Fluorescence
  • Transfection

Substances

  • Benzylidene Compounds
  • Receptors, G-Protein-Coupled
  • Receptors, Opioid, delta
  • Recombinant Proteins
  • 7-benzylidenenaltrexone
  • Naltrexone
  • Enkephalin, D-Penicillamine (2,5)-