Molecular Neurobiology

, Volume 56, Issue 10, pp 6756–6769 | Cite as

Biased G Protein-Independent Signaling of Dopamine D1-D3 Receptor Heteromers in the Nucleus Accumbens

  • Xavier Guitart
  • Estefanía Moreno
  • William Rea
  • Marta Sánchez-Soto
  • Ning-Sheng Cai
  • César Quiroz
  • Vivek Kumar
  • Liam Bourque
  • Antoni Cortés
  • Enric I. Canela
  • Christopher Bishop
  • Amy H. Newman
  • Vicent CasadóEmail author
  • Sergi FerréEmail author


Several studies found in vitro evidence for heteromerization of dopamine D1 receptors (D1R) and D3 receptors (D3R), and it has been postulated that functional D1R-D3R heteromers that are normally present in the ventral striatum mediate synergistic locomotor-activating effects of D1R and D3R agonists in rodents. Based also on results obtained in vitro, with mammalian transfected cells, it has been hypothesized that those behavioral effects depend on a D1R-D3R heteromer-mediated G protein-independent signaling. Here, we demonstrate the presence on D1R-D3R heteromers in the mouse ventral striatum by using a synthetic peptide that selectively destabilizes D1R-D3R heteromers. Parallel locomotor activity and ex vivo experiments in reserpinized mice and in vitro experiments in D1R-D3R mammalian transfected cells were performed to dissect the signaling mechanisms of D1R-D3R heteromers. Co-administration of D1R and D3R agonists in reserpinized mice produced synergistic locomotor activation and a selective synergistic AKT phosphorylation in the most ventromedial region of the striatum in the shell of the nucleus accumbens. Application of the destabilizing peptide in transfected cells and in the shell of the nucleus accumbens allowed demonstrating that both in vitro and in vivo co-activation of D3R induces a switch from G protein-dependent to G protein-independent D1R-mediated signaling determined by D1R-D3R heteromerization. The results therefore demonstrate that a biased G protein-independent signaling of D1R-D3R heteromers localized in the shell of the nucleus accumbens mediate the locomotor synergistic effects of D1R and D3R agonists in reserpinized mice.


GPCR heteromers Dopamine D1 receptor Dopamine D3 receptor Reserpine Functional selectivity 


Funding Information

This work is supported by the intramural funds of the National Institute on Drug Abuse, “Ministerio de Economía y Competitividad” and European Regional Development Funds of the European Union (Grants SAF2014-54840-R and SAF2017-87629-R), “Fundació La Marató de TV3” (Grant 20140610), and “Generalitat de Catalunya” (Grant 2017-SGR-1497).


  1. 1.
    Schwartz JC, Diaz J, Bordet R, Griffon N, Perachon S, Pilon C, Ridray S, Sokoloff P (1998) Functional implications of multiple dopamine receptor subtypes: the D1/D3 receptor coexistence. Brain Res Rev 26:236–242CrossRefGoogle Scholar
  2. 2.
    Sokoloff P, Giros B, Martres MP, Bouthenet ML, Schwartz JC (1990) Molecular cloning and characterization of a novel dopamine receptor (D3) as a target for neuroleptics. Nature 347:146–151CrossRefGoogle Scholar
  3. 3.
    Ridray S, Griffon N, Mignon V, Souil E, Carboni S, Diaz J, Schwartz JC, Sokoloff P (1998) Coexpression of dopamine D1 and D3 receptors in islands of Calleja and shell of nucleus accumbens of the rat: opposite and synergistic functional interactions. Eur J Neurosci 10:1676–1686CrossRefGoogle Scholar
  4. 4.
    Fiorentini C, Busi C, Gorruso E, Gotti C, Spano P, Missale C (2008) Reciprocal regulation of dopamine D1 and D3 receptor function and trafficking by heterodimerization. Mol Pharmacol 74:59–69CrossRefGoogle Scholar
  5. 5.
    Guitart X, Navarro G, Moreno E, Yano H, Cai NS, Sánchez-Soto M, Kumar-Barodia S, Naidu YT et al (2014) Functional selectivity of allosteric interactions within G protein-coupled receptor oligomers: the dopamine D1-D3 receptor heterotetramer. Mol Pharmacol 86:417–429CrossRefGoogle Scholar
  6. 6.
    Marcellino D, Ferré S, Casadó V, Cortés A, Le Foll B, Mazzola C, Drago F, Saur O et al (2008) Identification of dopamine D1-D3 receptor heteromers. Indications for a role of synergistic D1-D3 receptor interactions in the striatum. J Biol Chem 283:26016–26025CrossRefGoogle Scholar
  7. 7.
    Farré D, Muñoz A, Moreno E, Reyes-Resina I, Canet-Pons J, Dopeso-Reyes IG, Rico AJ, Lluís C et al (2015) Stronger dopamine D1 receptor-mediated neurotransmission in dyskinesia. Mol Neurobiol 52:1408–1420CrossRefGoogle Scholar
  8. 8.
    Ferré S, Lluis C, Lanciego JL, Franco R (2010) Prime time for G-protein-coupled receptor heteromers as therapeutic targets for CNS disorders: the dopamine D1-D3 receptor heteromer. CNS Neurol Disord Drug Targets 9:596–600CrossRefGoogle Scholar
  9. 9.
    Lanza K, Meadows SM, Chambers NE, Nuss E, Deak MM, Ferré S, Bishop C (2018) Behavioral and cellular dopamine D(1) and D(3) receptor-mediated synergy: implications for L-DOPA-induced dyskinesia. Neuropharmacology 138:304–314CrossRefGoogle Scholar
  10. 10.
    Ferré S, Giménez-Llort L, Artigas F, Martínez E (1994) Motor activation in short-and long-term reserpinized mice: role of N-methyl-D-aspartate, dopamine D1 and dopamine D2 receptors. Eur J Pharmacol 255:203–213CrossRefGoogle Scholar
  11. 11.
    Starr BS, Starr MS, Kilpatrick IC (1987) Behavioural role of dopamine D1 receptors in the reserpine-treated mouse. Neuroscience 22:179–188CrossRefGoogle Scholar
  12. 12.
    Missale C, Nash SR, Robinson SW, Jaber M, Caron MG (1998) Dopamine receptors: from structure to function. Physiol Rev 78:189–225CrossRefGoogle Scholar
  13. 13.
    Gilman AG (1987) G proteins: transducers of receptor-generated signals. Annu Rev Biochem 56:615–649CrossRefGoogle Scholar
  14. 14.
    Carriba P, Navarro G, Ciruela F, Ferré S, Casadó V, Agnati L, Cortés A, Mallol J et al (2008) Detection of heteromerization of more than two proteins by sequential BRET-FRET. Nat Methods 5:727–733CrossRefGoogle Scholar
  15. 15.
    Bonaventura J, Navarro G, Casadó-Anguera V, Azdad K, Rea W, Moreno E, Brugarolas M, Mallol J et al (2015) Allosteric interactions between agonists and antagonists within the adenosine A2A receptor-dopamine D2 receptor heterotetramer. Proc Natl Acad Sci U S A 112:E3609–E3618CrossRefGoogle Scholar
  16. 16.
    He SQ, Zhang ZN, Guan JS, Liu HR, Zhao B, Wang HB, Li Q, Yang H et al (2011) Facilitation of μ-opioid receptor activity by preventing δ-opioid receptor-mediated codegradation. Neuron 69:120–131CrossRefGoogle Scholar
  17. 17.
    Navarro G, Cordomí A, Casadó-Anguera V, Moreno E, Cai NS, Cortés A, Canela EI, Dessauer CW et al (2018) Evidence for functional pre-coupled complexes of receptor heteromers and adenylyl cyclase. Nat Commun 9:1242CrossRefGoogle Scholar
  18. 18.
    Rivera-Oliver M, Moreno E, Álvarez-Bagnarol Y, Ayala-Santiago C, Cruz-Reyes N, Molina-Castro GC, Clemens S, Canela EI et al (2018) Adenosine A(1)-dopamine D(1) receptor heteromers control the excitability of the spinal motoneuron. Mol Neurobiol 56:797–811Google Scholar
  19. 19.
    Schwarze SR, Ho A, Vocero-Akbani A, Dowdy SF (1999) In vivo protein transduction: delivery of a biologically active protein into the mouse. Science 285:1569–1572CrossRefGoogle Scholar
  20. 20.
    Kumar V, Bonifazi A, Ellenberger MP, Keck TM, Pommier E, Rais R, Slusher BS, Gardner E et al (2016) Highly selective dopamine D3 receptor (D3R) antagonists and partial agonists based on eticlopride and the D3R crystal structure: new leads for opioid dependence treatment. J Med Chem 59:7634–7650CrossRefGoogle Scholar
  21. 21.
    Yano H, Cai NS, Xu M, Verma RK, Rea W, Hoffman AF, Shi L, Javitch JA et al (2018) Gs- versus Golf-dependent functional selectivity mediated by the dopamine D(1) receptor. Nat Commun 9:486CrossRefGoogle Scholar
  22. 22.
    Andén NE, Grabowska-Andén M (1988) Stimulation of D1 dopamine receptors reveals direct effects of the preferential dopamine autoreceptor agonist B-HT 920 on postsynaptic dopamine receptors. Acta Physiol Scand 134:285–290CrossRefGoogle Scholar
  23. 23.
    Chen J, Rusnak M, Lombroso PJ, Sidhu A (2009) Dopamine promotes striatal neuronal apoptotic death via ERK signaling cascades. Eur J Neurosci 29:287–306CrossRefGoogle Scholar
  24. 24.
    Robinson SW, Jarvie KR, Caron MG (1994) High affinity agonist binding to the dopamine D3 receptor: chimeric receptors delineate a role for intracellular domains. Mol Pharmacol 46:352–356Google Scholar
  25. 25.
    Keck TM, John WS, Czoty PW, Nader MA, Newman AH (2015) Identifying medication targets for psychostimulant addiction: Unraveling the dopamine D3 receptor hypothesis. J Med Chem 58:5361–5380CrossRefGoogle Scholar
  26. 26.
    Beaulieu JM, Tirotta E, Sotnikova TD, Masri B, Salahpour A, Gainetdinov RR, Borrelli E, Caron MG (2007) Regulation of Akt signaling by D2 and D3 dopamine receptors in vivo. J Neurosci 27:881–885CrossRefGoogle Scholar
  27. 27.
    Salles MJ, Hervé D, Rivet JM, Longueville S, Millan MJ, Girault JA, Mannoury la Cour C (2013) Transient and rapid activation of Akt/GSK-3β and mTORC1 signaling by D3 dopamine receptor stimulation in dorsal striatum and nucleus accumbens. J Neurochem 125:532–544CrossRefGoogle Scholar
  28. 28.
    Ferré S (2015) The GPCR heterotetramer: challenging classical pharmacology. Trends Pharmacol Sci 36:145–152CrossRefGoogle Scholar
  29. 29.
    Costa-Neto CM, Parreiras-E-Silva LT, Bouvier M (2016) Pluridimensional view of biased agonism. Mol Pharmacol 90:587–595CrossRefGoogle Scholar
  30. 30.
    Reiter E, Ahn S, Shukla AK, Lefkowitz RJ (2012) Molecular mechanism of β-arrestin-biased agonism at seven-transmembrane receptors. Annu Rev Pharmacol Toxicol 52:179–197CrossRefGoogle Scholar
  31. 31.
    Urs NM, Peterson SM, Caron MG (2017) New concepts in dopamine D(2) receptor biased signaling and implications for schizophrenia therapy. Biol Psychiatry 81:78–85CrossRefGoogle Scholar
  32. 32.
    Violin JD, Crombie AL, Soergel DG, Lark MW (2014) Biased ligands at G-protein-coupled receptors: promise and progress. Trends Pharmacol Sci 35:308–316CrossRefGoogle Scholar
  33. 33.
    Rozenfeld R, Devi LA (2007) Receptor heterodimerization leads to a switch in signaling: beta-arrestin2-mediated ERK activation by mu-delta opioid receptor heterodimers. FASEB J 21:2455–2465CrossRefGoogle Scholar
  34. 34.
    Sahlholm K, Gómez-Soler M, Valle-León M, López-Cano M, Taura JJ, Ciruela F, Fernández-Dueñas V (2018) Antipsychotic-like efficacy of dopamine D(2) receptor-biased ligands is dependent on adenosine A(2A) receptor expression. Mol Neurobiol 55:4952–4958CrossRefGoogle Scholar
  35. 35.
    Gerfen CR, Miyachi S, Paletzki R, Brown P (2002) D1 dopamine receptor supersensitivity in the dopamine-depleted striatum results from a switch in the regulation of ERK1/2/MAP kinase. J Neurosci 22:5042–5054CrossRefGoogle Scholar
  36. 36.
    Prieto GA, Perez-Burgos A, Palomero-Rivero M, Galarraga E, Drucker-Colin R, Bargas J (2011) Upregulation of D2-class signaling in dopamine-denervated striatum is in part mediated by D3 receptors acting on Ca V 2.1 channels via PIP2 depletion. J Neurophysiol 105:2260–2274CrossRefGoogle Scholar
  37. 37.
    Bordet R, Ridray S, Carboni S, Diaz J, Sokoloff P, Schwartz JC (1997) Induction of dopamine D3 receptor expression as a mechanism of behavioral sensitization to levodopa. Proc Natl Acad Sci U S A 94:3363–3367CrossRefGoogle Scholar
  38. 38.
    Solís O, Garcia-Montes JR, González-Granillo A, Xu M, Moratalla R (2017) Dopamine D3 receptor modulates l-DOPA-induced dyskinesia by targeting D1 receptor-mediated striatal signaling. Cereb Cortex 27:435–446Google Scholar
  39. 39.
    Bychkov E, Ahmed MR, Dalby KN, Gurevich EV (2007) Dopamine depletion and subsequent treatment with L-DOPA, but not the long-lived dopamine agonist pergolide, enhances activity of the Akt pathway in the rat striatum. J Neurochem 102:699–711CrossRefGoogle Scholar
  40. 40.
    Morissette M, Samadi P, Hadj Tahar A, Bélanger N, Di Paolo T (2010) Striatal Akt/GSK3 signaling pathway in the development of L-Dopa-induced dyskinesias in MPTP monkeys. Prog Neuro-Psychopharmacol Biol Psychiatry 34:446–454CrossRefGoogle Scholar
  41. 41.
    Staley JK, Mash DC (1996) Adaptive increase in D3 dopamine receptors in the brain reward circuits of human cocaine fatalities. J Neurosci 16:6100–6106CrossRefGoogle Scholar

Copyright information

© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2019

Authors and Affiliations

  • Xavier Guitart
    • 1
  • Estefanía Moreno
    • 2
  • William Rea
    • 1
  • Marta Sánchez-Soto
    • 1
  • Ning-Sheng Cai
    • 1
  • César Quiroz
    • 1
  • Vivek Kumar
    • 3
  • Liam Bourque
    • 1
  • Antoni Cortés
    • 2
  • Enric I. Canela
    • 2
  • Christopher Bishop
    • 4
  • Amy H. Newman
    • 3
  • Vicent Casadó
    • 2
    Email author
  • Sergi Ferré
    • 1
    Email author
  1. 1.Integrative Neurobiology SectionNational Institute on Drug Abuse, Intramural Research Program, National Institutes of HealthBaltimoreUSA
  2. 2.Department of Biochemistry and Molecular Biomedicine of the Faculty of Biology and Institute of Biomedicine of the University of Barcelona and Center for Biomedical Research in Neurodegenerative Diseases NetworkBarcelonaSpain
  3. 3.Medicinal Chemistry SectionNational Institute on Drug Abuse, Intramural Research Program, National Institutes of HealthBaltimoreUSA
  4. 4.Behavioral Neuroscience Program, Department of PsychologyBinghamton UniversityBinghamtonUSA

Personalised recommendations