title: BRET approaches to characterize dopamine and TAAR1 receptor pharmacology and signaling.
abstract: 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.
authors.affiliation: Department of Neuroscience and Brain Technologies, Italian Institute of Technology, Genoa, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Espinoza
authors.forename: Bernard
authors.initials: B
authors.lastname: Masri
authors.forename: Ali
authors.initials: A
authors.lastname: Salahpour
authors.forename: Raul R
authors.initials: RR
authors.lastname: Gainetdinov
mesh_terms: D019390:Arrestins / Q000378:metabolism; D060807:Bioluminescence Resonance Energy Transfer Techniques / Q000379:methods*; D015374:Biosensing Techniques; D000242:Cyclic AMP / Q000378:metabolism; D004298:Dopamine / Q000378:metabolism*; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*; D014162:Transfection; D000071557:beta-Arrestins; D000095383:Trace Amine-Associated Receptors
chemical_list: D019390:Arrestins; D000242:Cyclic AMP; D004298:Dopamine; D020662:Guanine Nucleotide Exchange Factors; D043562:Receptors, G-Protein-Coupled; D000071557:beta-Arrestins; D000095383:Trace Amine-Associated Receptors