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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
Article

Abstract

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.

Keywords

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

Notes

Acknowledgements

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).

Funding

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.

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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

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