TY - CHAP
T1 - Detection and quantitative analysis of dynamic GPCRs interactions using flow cytometry-based FRET
AU - Chruścicka, Barbara
AU - Wallace Fitzsimons, Shauna E.
AU - Druelle, Clémentine M.
AU - Dinan, Timothy G.
AU - Schellekens, Harriët
N1 - Publisher Copyright:
© Springer Science+Business Media, LLC, part of Springer Nature 2018.
PY - 2018
Y1 - 2018
N2 - Heterodimerization of specific G protein-coupled receptor (GPCR) protomers is associated with increased receptor signaling diversity and exhibits unique biochemical, functional, and pharmacological properties. Evidence for the formation of heteroreceptor complexes has been demonstrated in vitro using cellular models and biochemical assays and ex vivo using brain slices and primary cell cultures. Since mechanisms that lead to brain pathologies such as depression, anxiety, addiction, and schizophrenia involve GPCR signaling, the distinct pharmacological profiles of GPCR assemblies may serve as new target for the development of novel therapeutic strategies with enhanced specificity. Therefore, development and standardization of novel methods for detection and analysis of dimer pairs both in recombinant systems and in native tissue is warranted. This chapter describes a step-by-step protocol for detecting and quantifying dynamic receptor–receptor interactions in living cells using flow cytometry-based fluorescence (Förster) resonance energy transfer (fcFRET). This method has significant potential to identify novel GPCR dimers within the central nervous system while simultaneously allowing analysis of the dynamic nature of these receptor interactions, which is poised to contribute significantly to the field of GPCR neuropsychopharmacology across brain diseases.
AB - Heterodimerization of specific G protein-coupled receptor (GPCR) protomers is associated with increased receptor signaling diversity and exhibits unique biochemical, functional, and pharmacological properties. Evidence for the formation of heteroreceptor complexes has been demonstrated in vitro using cellular models and biochemical assays and ex vivo using brain slices and primary cell cultures. Since mechanisms that lead to brain pathologies such as depression, anxiety, addiction, and schizophrenia involve GPCR signaling, the distinct pharmacological profiles of GPCR assemblies may serve as new target for the development of novel therapeutic strategies with enhanced specificity. Therefore, development and standardization of novel methods for detection and analysis of dimer pairs both in recombinant systems and in native tissue is warranted. This chapter describes a step-by-step protocol for detecting and quantifying dynamic receptor–receptor interactions in living cells using flow cytometry-based fluorescence (Förster) resonance energy transfer (fcFRET). This method has significant potential to identify novel GPCR dimers within the central nervous system while simultaneously allowing analysis of the dynamic nature of these receptor interactions, which is poised to contribute significantly to the field of GPCR neuropsychopharmacology across brain diseases.
KW - Dimerization
KW - Flow cytometry
KW - Fluorescent resonance energy transfer
KW - G protein-coupled receptors
KW - Heteroreceptor complexes
KW - Receptor–receptor interactions
UR - https://www.scopus.com/pages/publications/85051237281
U2 - 10.1007/978-1-4939-8576-0_14
DO - 10.1007/978-1-4939-8576-0_14
M3 - Chapter
AN - SCOPUS:85051237281
T3 - Neuromethods
SP - 223
EP - 238
BT - Neuromethods
PB - Humana Press Inc.
ER -