Molecular Neurobiology

, Volume 56, Issue 7, pp 4778–4785 | Cite as

Revisiting the Functional Role of Dopamine D4 Receptor Gene Polymorphisms: Heteromerization-Dependent Gain of Function of the D4.7 Receptor Variant

  • Marta Sánchez-Soto
  • Hideaki Yano
  • Ning-Sheng Cai
  • Verònica Casadó-Anguera
  • Estefanía Moreno
  • Vicent Casadó
  • Sergi FerréEmail author


The two most common polymorphisms of the human DRD4 gene encode a dopamine D4 receptor (D4R) with four or seven repeats of a proline-rich sequence of 16 amino acids (D4.4R or D4.7R). Although the seven-repeat polymorphism has been repeatedly associated with attention-deficit hyperactivity disorder and substance use disorders, the differential functional properties between D4.4R and D4.7R remained enigmatic until recent electrophysiological and optogenetic-microdialysis experiments indicated a gain of function of D4.7R. Since no clear differences in the biochemical properties of individual D4.4R and D4.7R have been reported, it was previously suggested that those differences emerge upon heteromerization with dopamine D2 receptor (D2R), which co-localizes with D4R in the brain. However, contrary to a gain of function, experiments in mammalian transfected cells suggested that heteromerization with D2R results in lower MAPK signaling by D4.7R as compared to D4.4R. In the present study, we readdressed the question of functional differences of D4.4R and D4.7R forming homomers or heteromers with the short isoform of D2R (D2SR), using a functional bioluminescence resonance energy transfer (BRET) assay that allows the measurement of ligand-induced changes in the interaction between G protein-coupled receptors (GPCRs) forming homomers or heteromers with their cognate G protein. Significant functional and pharmacological differences between D4.4R and D4.7R were only evident upon heteromerization with the short isoform of D2R (D2SR). The most dramatic finding was a significant increase and decrease in the constitutive activity of D2S upon heteromerization with D4.7R and D4.4R, respectively, providing the first clear mechanism for a functional difference between both products of polymorphic variants and for a gain of function of the D4.7R.


Dopamine D4 receptor Dopamine D2 receptor Gene polymorphisms G protein-coupled receptor heteromers Constitutive activity Bioluminescence resonance energy transfer 



We thank Dr. J. A. Javitch (Columbia University, New York) for kindly providing some of the DNA constructs (D2SR-nRLuc, D2SR-cRLuc, and Gαi1-YFP).


This work is supported by the intramural funds of the National Institute on Drug Abuse, a grant from the Spanish “Ministerio de Economía y Competitividad,” and the European Regional Development Funds of the European Union (SAF2014-54840-R).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.


  1. 1.
    LaHoste GJ, Swanson JM, Wigal SB, Glabe C, Wigal T, King N, Kennedy JL (1996) Dopamine D4 receptor gene polymorphism is associated with attention deficit hyperactivity disorder. Mol Psychiatry 1:121–124PubMedGoogle Scholar
  2. 2.
    Chang FM, Kidd JR, Livak KJ, Pakstis AJ, Kidd KK (1996) The world-wide distribution of allele frequencies at the human dopamine D4 receptor locus. Hum Genet 98:91–101CrossRefGoogle Scholar
  3. 3.
    Wang E, Ding YC, Flodman P, Kidd JR, Kidd KK, Grady DL, Ryder OA, Spence MA (2004) The genetic architecture of selection at the human dopamine receptor D4 (DRD4) gene locus. Am J Hum Genet 74:931–944CrossRefGoogle Scholar
  4. 4.
    Faraone SV, Perlis RH, Doyle AE, Smoller JW, Goralnick JJ, Holmgren MA, Sklar P (2005) Molecular genetics of attention-deficit/hyperactivity disorder. Biol Psychiatry 57:1313–1323CrossRefGoogle Scholar
  5. 5.
    Li D, Sham PC, Owen MJ, He L (2006) Meta-analysis shows significant association between dopamine system genes and attention deficit hyperactivity disorder (ADHD). Hum Mol Genet 15:2276–2284CrossRefGoogle Scholar
  6. 6.
    Gizer IR, Ficks C, Waldman ID (2009) Candidate gene studies of ADHD: a meta-analytic review. Hum Genet 126:51–90CrossRefGoogle Scholar
  7. 7.
    Belcher AM, Volkow ND, Moeller FG, Ferré S (2014) Personality traits and vulnerability or resilience to substance use disorders. Trends Cogn Sci 18:211–217CrossRefGoogle Scholar
  8. 8.
    Zhong P, Liu W, Yan Z (2016) Aberrant regulation of synchronous network activity by the attention-deficit/hyperactivity disorder-associated human dopamine D4 receptor variant D4.7 in the prefrontal cortex. J Physiol 594:135–147CrossRefGoogle Scholar
  9. 9.
    Bonaventura J, Quiroz C, Cai NS, Rubinstein M, Tanda G, Ferré S (2017) Key role of the dopamine D(4) receptor in the modulation of corticostriatal glutamatergic neurotransmission. Sci Adv 3:e1601631CrossRefGoogle Scholar
  10. 10.
    González S, Rangel-Barajas C, Peper M, Lorenzo R, Moreno E, Ciruela F, Borycz J, Ortiz J et al (2012) Dopamine D4 receptor, but not the ADHD-associated D4.7 variant, forms functional heteromers with the dopamine D2S receptor in the brain. Mol Psychiatry 17:650–662CrossRefGoogle Scholar
  11. 11.
    Asghari V, Sanyal S, Buchwaldt S, Paterson A, Jovanovic V, Van Tol HH (1995) Modulation of intracellular cyclic AMP levels by different human dopamine D4 receptor variants. J Neurochem 65:1157–1165CrossRefGoogle Scholar
  12. 12.
    Sánchez-Soto M, Bonifazi A, Cai N-S, Ellenberger MP, Newman AH, Ferré S, Yano H (2016) Evidence for noncanonical neurotransmitter activation: norepinephrine as a dopamine D2-like receptor agonist. Mol Pharmacol 89:457–466CrossRefGoogle Scholar
  13. 13.
    Yepes G, Guitart X, Rea W, Richard PA, Earley CJ, Quiroz C, Ferré S (2017) Targeting hypersensitive corticostriatal terminals in restless legs syndrome. Ann Neurol 82:951–960CrossRefGoogle Scholar
  14. 14.
    Borroto-Escuela DO, Van Craenenbroeck K, Romero-Fernandez W, Guidolin D, Woods AS, Rivera A, Haegeman G, Agnati LF et al (2011) Dopamine D2 and D4 receptor heteromerization and its allosteric receptor-receptor interactions. Biochem Biophys Res Commun 404:928–934CrossRefGoogle Scholar
  15. 15.
    Urizar E, Yano H, Kolster R, Galés C, Lambert N, Javitch JA (2011) CODA-RET reveals functional selectivity as a result of GPCR heteromerization. Nat Chem Biol 7:624–630CrossRefGoogle Scholar
  16. 16.
    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
  17. 17.
    Ferré S, Casadó V, Devi LA, Filizola M, Jockers R, Lohse MJ, Milligan G, Pin J-P et al (2014) G protein-coupled receptor oligomerization revisited: functional and pharmacological perspectives. Pharmacol Rev 66:413–434CrossRefGoogle Scholar
  18. 18.
    Patel S, Freedman S, Chapman KL, Emms F, Fletcher AE, Knowles M, Marwood R, Mcallister G et al (1997) Biological profile of L-745,870, a selective antagonist with high affinity for the dopamine D4 receptor. J Pharmacol Exp Ther 283:636–647PubMedGoogle Scholar
  19. 19.
    Nilsson CL, Ekman A, Hellstrand M, Eriksson E (1996) Inverse agonism at dopamine D2 receptors. Haloperidol-induced prolactin release from GH4C1 cells transfected with the human D2 receptor is antagonized by R(−)-n-propylnorapomorphine, raclopride, and phenoxybenzamine. Neuropsychopharmacology 15:53–61CrossRefGoogle Scholar

Copyright information

© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2018

Authors and Affiliations

  1. 1.Integrative Neurobiology Section, National Institute on Drug Abuse, Intramural Research ProgramNational Institutes of HealthBaltimoreUSA
  2. 2.Department of Biochemistry and Molecular Biomedicine, Faculty of Biology, Institute of Biomedicine of the University of BarcelonaUniversity of Barcelona, and Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED)BarcelonaSpain

Personalised recommendations