DNA microarray analysis of cannabinoid signaling in mouse brain in vivo.
Sophie Parmentier-Batteur, Kunlin Jin, Lin Xie, Xiao Ou Mao, David A Greenberg
Molecular Pharmacology October 2002 DOI: 10.1124/mol.62.4.828 (opens in new tab)
Study at a glance
AI-extracted from the abstract| Characteristics | Observational study Peer reviewed |
|---|---|
| Population | Mouse forebrain |
| Interventions | Delta(9)-tetrahydrocannabinol (Delta(9)-THC) R(+)-WIN 55 212-2 |
| Duration | 12 h after a single dose |
| Measures | cDNA microarrays, Western blotting, immunohistochemistry |
| Key findings | A single dose of Delta(9)-THC or R(+)-WIN 55,212-2 altered forebrain expression of 65 and 41 genes respectively, with 20 genes changed by both. Two shared genes, MRG1 and glucokinase, showed roughly 2-fold protein increases, with MRG1 in cerebral blood vessels and GK in hypothalamic neurons, suggesting they are cannabinoid-regulated genes possibly involved in vascular and hypothalamic cannabinoid effects. |
Abstract
To identify novel genes involved in cannabinoid receptor-mediated signaling, we used cDNA microarrays to detect changes in mRNA expression in the forebrains of mice 12 h after they were given a single intraperitoneal dose of the naturally-occurring Cannabis sativa alkaloid Delta(9)-tetrahydrocannabinol (Delta(9)-THC) or the synthetic cannabinoid receptor agonist (R)-(+)-2,3-dihydro-5-methyl-3-[(morpholinyl)methyl] pyrrolo[1,2,3-de]-1,4-benzoxazin-yl-1-naphtalenylmethanone mesylate [R(+)-WIN 55,212-2]. Of approximately 11,000 genes from a mouse brain cDNA library that were probed, 65 showed altered (increased or decreased at least 2-fold) expression after exposure to Delta(9)-THC, 41 after exposure to R(+)-WIN 55,212-2, and 20 genes after exposure to both drugs. Genes affected similarly by Delta(9)-THC and R(+)-WIN 55,212-2 were considered likely to reflect cannabinoid receptor activation, and expression of the protein products of two such genes not previously implicated in cannabinoid signaling-melanocyte-specific gene-related gene 1 (MRG1) and hexokinase 4 (glucokinase, GK)-was measured by Western blotting and immunohistochemistry. Western blots showed approximately 2-fold increases in the levels of both proteins in mouse forebrain. Immunohistochemistry revealed preferential localization of MRG1 to cerebral blood vessels and of GK to hypothalamic neurons. These findings suggest that MRG1 and GK are cannabinoid-regulated genes and that they may be involved in the vascular and hypothalamic effects of cannabinoids, respectively.