A G alpha sub(s) DREADD Mouse for Selective Modulation of cAMP Production in Striatopallidal Neurons

• Published in: Neuropsychopharmacology (New York, N.Y.) N.Y.), 2013-04, Vol.38 (5), p.854-862
• Main Authors: Farrell, Martilias S, Pei, Ying, Wan, Yehong, Yadav, Prem N, Daigle, Tanya L, Urban, Daniel J, Lee, Hyeong-Min, Sciaky, Noah, Simmons, Arkeen, Nonneman, Randal J, Huang, Xi-Ping, Hufeisen, Sandy J, Guettier, Jean-Marc, Moy, Sheryl S, Wess, Juergen, Caron, Marc G, Calakos, Nicole, Roth, Bryan L
• Format: Article
• Language: English
• Subjects: alpha -Amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors; Amphetamine; Bacterial artificial chromosomes; Cyclic AMP; DARPP-32 protein; G protein-coupled receptors; Glutamic acid receptors; Locomotor activity; N-Methyl-D-aspartic acid receptors; Neostriatum; Novelty; Nucleus accumbens; Phosphorylation; spiny neurons; Transgenic mice
• ISSN: 0893-133X
• DOI: 10.1038/npp.2012.251

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 G alpha sub(olf) 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 G alpha sub(s)-coupled GPCRs. We then tested whether CNO administration would produce behavioral responses associated with striatopallidal G sub(s) 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 G alpha sub(s) signaling in specific neuronal populations in vivo.This unique tool provides a new resource for elucidating the roles of striatopallidal MSN G alpha sub(s) signaling in other neurobehavioral contexts.