Metabotropic glutamate type 5, dopamine D2 and adenosine A2a receptors form higher-order oligomers in living cells

J Neurochem. 2009 Jun;109(5):1497-507. doi: 10.1111/j.1471-4159.2009.06078.x. Epub 2009 Mar 30.

Abstract

G protein-coupled receptors are known to form homo- and heteromers at the plasma membrane, but the stoichiometry of these receptor oligomers are relatively unknown. Here, by using bimolecular fluorescence complementation, we visualized for the first time the occurrence of heterodimers of metabotropic glutamate mGlu(5) receptors (mGlu(5)R) and dopamine D(2) receptors (D(2)R) in living cells. Furthermore, the combination of bimolecular fluorescence complementation and bioluminescence resonance energy transfer techniques, as well as the sequential resonance energy transfer technique, allowed us to detect the occurrence receptor oligomers containing more than two protomers, mGlu(5)R, D(2)R and adenosine A(2A) receptor (A(2A)R). Interestingly, by using high-resolution immunoelectron microscopy we could confirm that the three receptors co-distribute within the extrasynaptic plasma membrane of the same dendritic spines of asymmetrical, putative glutamatergic, striatal synapses. Also, co-immunoprecipitation experiments in native tissue demonstrated the existence of an association of mGlu(5)R, D(2)R and A(2A)R in rat striatum homogenates. Overall, these results provide new insights into the molecular composition of G protein-coupled receptor oligomers in general and the mGlu(5)R/D(2)R/A(2A)R oligomer in particular, a receptor oligomer that might constitute an important target for the treatment of some neuropsychiatric disorders.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins
  • Cell Line, Transformed / metabolism
  • Cell Line, Transformed / ultrastructure
  • Corpus Striatum / cytology
  • Dimerization
  • Fluorescence Resonance Energy Transfer / methods
  • Humans
  • Immunoprecipitation
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Microscopy, Electron, Transmission / methods
  • Models, Molecular
  • Rats
  • Receptor, Adenosine A2A / chemistry
  • Receptor, Adenosine A2A / genetics
  • Receptor, Adenosine A2A / metabolism*
  • Receptor, Adenosine A2A / ultrastructure
  • Receptor, Metabotropic Glutamate 5
  • Receptors, Dopamine D2 / chemistry
  • Receptors, Dopamine D2 / genetics
  • Receptors, Dopamine D2 / metabolism*
  • Receptors, Dopamine D2 / ultrastructure
  • Receptors, Metabotropic Glutamate / chemistry
  • Receptors, Metabotropic Glutamate / genetics
  • Receptors, Metabotropic Glutamate / metabolism*
  • Receptors, Metabotropic Glutamate / ultrastructure
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Transfection

Substances

  • Bacterial Proteins
  • Luminescent Proteins
  • Receptor, Adenosine A2A
  • Receptor, Metabotropic Glutamate 5
  • Receptors, Dopamine D2
  • Receptors, Metabotropic Glutamate
  • Recombinant Fusion Proteins
  • yellow fluorescent protein, Bacteria