A Gαs DREADD mouse for selective modulation of cAMP production in striatopallidal neurons

Neuropsychopharmacology. 2013 Apr;38(5):854-62. doi: 10.1038/npp.2012.251. Epub 2012 Dec 5.

Abstract

Here, we describe a newly generated transgenic mouse in which the Gs DREADD (rM3Ds), an engineered G protein-coupled receptor, is selectively expressed in striatopallidal medium spiny neurons (MSNs). We first show that in vitro, rM3Ds can couple to Gαolf and induce cAMP accumulation in cultured neurons and HEK-T cells. The rM3Ds was then selectively and stably expressed in striatopallidal neurons by creating a transgenic mouse in which an adenosine2A (adora2a) receptor-containing bacterial artificial chromosome was employed to drive rM3Ds expression. In the adora2A-rM3Ds mouse, activation of rM3Ds by clozapine-N-oxide (CNO) induces DARPP-32 phosphorylation, consistent with the known consequence of activation of endogenous striatal Gαs-coupled GPCRs. We then tested whether CNO administration would produce behavioral responses associated with striatopallidal Gs signaling and in this regard CNO dose-dependently decreases spontaneous locomotor activity and inhibits novelty induced locomotor activity. Last, we show that CNO prevented behavioral sensitization to amphetamine and increased AMPAR/NMDAR ratios in transgene-expressing neurons of the nucleus accumbens shell. These studies demonstrate the utility of adora2a-rM3Ds transgenic mice for the selective and noninvasive modulation of Gαs signaling in specific neuronal populations in vivo.This unique tool provides a new resource for elucidating the roles of striatopallidal MSN Gαs signaling in other neurobehavioral contexts.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adrenergic Uptake Inhibitors / pharmacology
  • Amphetamine / pharmacology
  • Animals
  • Animals, Newborn
  • Clozapine / analogs & derivatives
  • Clozapine / pharmacology
  • Cocaine / pharmacology
  • Corpus Striatum / cytology*
  • Cyclic AMP / metabolism*
  • Dopamine Uptake Inhibitors / pharmacology
  • Dopamine and cAMP-Regulated Phosphoprotein 32 / metabolism
  • GTP-Binding Protein alpha Subunits, Gs / genetics*
  • Green Fluorescent Proteins / genetics
  • Locomotion / drug effects
  • Locomotion / genetics
  • Luminescent Proteins / genetics
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Neurons / metabolism*
  • Phosphorylation / drug effects
  • Phosphorylation / genetics
  • Receptor, Adenosine A2A / genetics
  • Receptors, Dopamine D1 / genetics
  • Receptors, Dopamine D2 / genetics
  • Receptors, G-Protein-Coupled / genetics*
  • Receptors, Muscarinic / genetics*
  • Red Fluorescent Protein

Substances

  • Adrenergic Uptake Inhibitors
  • Dopamine Uptake Inhibitors
  • Dopamine and cAMP-Regulated Phosphoprotein 32
  • Luminescent Proteins
  • Receptor, Adenosine A2A
  • Receptors, Dopamine D1
  • Receptors, Dopamine D2
  • Receptors, G-Protein-Coupled
  • Receptors, Muscarinic
  • Green Fluorescent Proteins
  • Amphetamine
  • Cyclic AMP
  • GTP-Binding Protein alpha Subunits, Gs
  • Cocaine
  • Clozapine
  • clozapine N-oxide