Relative Role of Akt, ERK and CREB in Alcohol-Induced Microglia P2X4R Receptor Expression

• Published in: Alcohol and alcoholism (Oxford) 2016-11, Vol.51 (6), p.647-654
• Main Authors: Gofman, Larisa, Fernandes, Nicole C, Potula, Raghava
• Format: Article
• Language: English
• Subjects: Blotting, Western; Brain-Derived Neurotrophic Factor - physiology; Cyclic AMP Response Element-Binding Protein - physiology; Ethanol - pharmacology; Gene Expression Regulation - drug effects; Gene Expression Regulation - physiology; Gene Knockdown Techniques; Human Embryonic Stem Cells; Humans; MAP Kinase Signaling System - physiology; Microglia - drug effects; Microglia - physiology; Oncogene Protein v-akt - physiology; Real-Time Polymerase Chain Reaction; Receptors, Purinergic P2X4 - metabolism; Receptors, Purinergic P2X4 - physiology; Signal Transduction - drug effects; Signal Transduction - physiology
• ISSN: 0735-0414; 1464-3502; 1464-3502
• DOI: 10.1093/alcalc/agw009

Previously we have demonstrated altered microglia P2X4R expression in response to alcohol and pharmacological blockade with a selective P2X4R antagonist can reverse the action, suggesting that P2X4R play a role in mediating alcohol-induced effects on microglia. In the present study, we investigated the underlying signaling mediators, which may play a role in modulating P2X4R expression in microglia cells in response to alcohol. Embryonic stem cell-derived microglia (ESdM) cells were used to investigate the potential mechanisms involved in the regulation of P2X4R in response to alcohol. Selective P2X4R antagonist and kinase inhibitors were used to further corroborate the signal transduction pathway through which alcohol modulates P2X4R expression in microglia. Alcohol (100 mM) suppressed phosphorylated AKT and ERK cascades in native ESdM cells. This alcohol-induced suppression was confirmed to be P2X4R-dependent through the use of a selective P2X4R antagonist and knockdown of P2XR4 by siRNA. Alcohol increased transcriptional activity of CREB. P2X4R antagonist blocked alcohol-induced effects on CREB, suggesting a P2X4R-mediated effect. These findings provide important clues to the underlying mechanism of purinoceptors in alcohol-induced microglia immune suppression.