Quantitative Proteomic Methodology Use and Development to Characterize Ethanol Modulation of Microglial Function

Microglia act as the frontline immune defense in the brain. Microglial responses can be either neurotoxic, through the release of reactive oxygen and nitrogen species and inflammatory cytokines, or neurotrophic. Microglial activation due to chronic ethanol exposure has been implicated in neuroinflammation. We use mass spectrometric metabolic labeling techniques to explore and quantify the microglial proteome in immortalized cell lines and in vivo enriched microglia. Our proteomic profiling and subsequent validation suggests that microglia do activate in response to ethanol exposure, but the activation falls short of the classical, or M1 state of inflammatory activation, as no downstream markers for reactive species nor inflammatory cytokines can be found. Additionally, proteomic profiling suggests a partial activation marked by increased cell engulfment and cell movement in addition to increased release of inf-gamma and tgf-beta