in the cell’s cytosol, likely in vesicles, before its concentration in the surrounding fluid increases progressively, consistent with neuronal release (Zou and Crews 2014). The importance of ethanol-induced release of HMGB1 and resulting TLR4 activation to ethanol-induced neurodegeneration and behavioral pathology was demonstrated in studies using cells and animals that no longer produced TLR4 (i.e., TLR4 knockout cells and mice). The experiments showed that knockout of TLR4 markedly blunted chronic–ethanol-induced neurodegeneration and induction of proinflammatory gene expression (Alfonso-Loeches et al. 2010; Blanco et al. 2005; Fernandez-Lizarbe et al. 2009; Pascual et al. 2011; Valles et al. 2004).
