Dopamine pp 107-122 | Cite as

BRET Approaches to Characterize Dopamine and TAAR1 Receptor Pharmacology and Signaling

  • Stefano Espinoza
  • Bernard Masri
  • Ali Salahpour
  • Raul R. GainetdinovEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 964)


It is evident that G protein-coupled receptors (GPCRs) such as D2 dopamine receptor and functionally related Trace Amine Associated Receptor 1 (TAAR1) can engage both in G protein-dependent (e.g., cAMP-mediated) and -independent β-arrestin-mediated signaling modalities. Both of these signaling events can be monitored in real-time and in live cells by using new biosensors based on a Bioluminescence Resonance Energy Transfer (BRET) approach. Here we discuss the practical applications of BRET to analyze dynamics of cAMP modulation via an EPAC biosensor as well as recruitment of β-arrestin2 to the D2 dopamine receptor. Combination of these approaches allows for a comparison of activity of pharmacological compounds on these signaling modalities as demonstrated for various antipsychotics as regard to D2 dopamine receptor. Furthermore, analysis of cAMP concentrations in cells expressing TAAR1 provides a simple high-throughput screening method to identify new ligands for this receptor. These BRET approaches could be applied for the characterization of pharmacology and signaling of variety of other GPCRs.

Key words

D2R TAAR1 cAMP β-arrestin2 BRET EPAC 3-Methoxytyramine β-Phenylethylamine 



Supported in part by research grants from the Michael J. Fox Foundation for Parkinson’s Research, Fondazione Compagnia di San Paolo (Torino, Italy) and research grant from F. Hoffmann-La Roche Ltd. (Basel, Switzerland) to Raul R. Gainetdinov. Bernard Masri was a recipient of a European Marie-Curie Outgoing International Fellowship (FP6—2005-Mobility-6). Ali Salahpour is supported by grants from Canadian Institute of Health Research (CIHR # 210296) and Natural Sciences and Engineering Council of Canada (NSERC # 386422).


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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Stefano Espinoza
    • 1
  • Bernard Masri
    • 2
  • Ali Salahpour
    • 3
  • Raul R. Gainetdinov
    • 1
    Email author
  1. 1.Department of Neuroscience and Brain TechnologiesItalian Institute of TechnologyGenoaItaly
  2. 2.Cancer Research Center of ToulouseINSERM U1037 - Université Paul Sabatier Toulouse III, CHU RangueilToulouseFrance
  3. 3.Department of Pharmacology and ToxicologyUniversity of TorontoTorontoCanada

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