Histone methyltransferase G9a diminishes expression of cannabinoid CB1 receptors in primary sensory neurons in neuropathic pain

Abstract

Type 1 cannabinoid receptors (CB₁Rs) are expressed in the dorsal root ganglion (DRG) and contribute to the analgesic effect of cannabinoids. However, the epigenetic mechanism regulating the expression of CB₁Rs in neuropathic pain is unknown. G9a (encoded by the <i>Ehmt2</i> gene), a histone 3 at lysine 9 methyltransferase, is a key chromatin regulator responsible for gene silencing. In this study, we determined G9a's role in regulating CB₁R expression in the DRG and in CB₁R-mediated analgesic effects in an animal model of neuropathic pain. We show that nerve injury profoundly reduced mRNA levels of CB₁Rs but increased the expression of CB₂ receptors in the rat DRG. ChIP results indicated increased enrichment of histone 3 at lysine 9 dimethylation, a G9a-catalyzed repressive histone mark, at the promoter regions of the CB₁R genes. G9a inhibition in nerve-injured rats not only up-regulated the CB₁R expression level in the DRG but also potentiated the analgesic effect of a CB₁R agonist on nerve injury-induced pain hypersensitivity. Furthermore, in mice lacking <i>Ehmt2</i> in DRG neurons, nerve injury failed to reduce CB₁R expression in the DRG and to decrease the analgesic effect of the CB₁R agonist. Moreover, nerve injury diminished the inhibitory effect of the CB₁R agonist on synaptic glutamate release from primary afferent nerves to spinal cord dorsal horn neurons in WT mice but not in mice lacking <i>Ehmt2</i> in DRG neurons. Our findings reveal that nerve injury diminishes the analgesic effect of CB₁R agonists through G9a-mediated CB₁R down-regulation in primary sensory neurons.