D(2)-Dopamine receptors target regulator of G protein signaling 9-2 to detergent-resistant membrane fractions

J Neurochem. 2012 Jan;120(1):56-69. doi: 10.1111/j.1471-4159.2011.07559.x. Epub 2011 Nov 24.

Abstract

Detergent-resistant membranes (DRM) are thought to contain structures such as lipid rafts that are involved in compartmentalizing cell membranes. We report that the majority of D(2)-dopamine receptors (D(2)R) expressed endogenously in mouse striatum or expressed in immortalized cell-lines is found in DRM. In addition, exogenous co-expression of D(2)R in a cell line shifted the expression of regulator of G protein signaling 9-2 (RGS9-2) into DRM. RGS9-2 is a protein that is highly enriched in the striatum and specifically regulates striatal D(2)R. In the striatum, RGS9-2 is mostly associated with DRMs but when expressed in cell lines, RGS9-2 is present in the soluble cytoplasmic fraction. In contrast, the majority of mu opioid receptors and delta opioid receptors are found in detergent-soluble membrane and there was no shift of RGS9-2 into DRM after co-expression of mu opioid receptor. These data suggest that the targeting of RGS9-2 to DRM in the striatum is mediated by D(2)R and that DRM is involved in the formation of a D(2)R signaling complex. D(2)R-mediated targeting of RGS9-2 to DRM was blocked by the deletion of the RGS9-2 DEP domain or by a point mutation that abolishes the GTPase accelerating protein function of RGS9-2.

Publication types

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

MeSH terms

  • Blotting, Western
  • Brain Chemistry
  • Cell Line
  • Cell Membrane / drug effects*
  • Corpus Striatum / metabolism
  • DNA, Complementary / genetics
  • Detergents / pharmacology*
  • GTP-Binding Protein beta Subunits / genetics
  • Humans
  • Immunohistochemistry
  • Membrane Proteins / chemistry
  • Mutation / genetics
  • Octoxynol
  • RGS Proteins / genetics
  • RGS Proteins / physiology*
  • Receptors, Dopamine D2 / drug effects*
  • Receptors, Dopamine D2 / genetics
  • Receptors, Opioid, delta / genetics
  • Receptors, Opioid, mu / genetics
  • Receptors, Opioid, mu / physiology
  • Signal Transduction / drug effects*
  • Solubility
  • Transfection

Substances

  • DNA, Complementary
  • Detergents
  • GNB5 protein, human
  • GTP-Binding Protein beta Subunits
  • Membrane Proteins
  • RGS Proteins
  • Receptors, Dopamine D2
  • Receptors, Opioid, delta
  • Receptors, Opioid, mu
  • regulator of g-protein signaling 9
  • Octoxynol