Acute ethanol exposure disrupts actin cytoskeleton and generates reactive oxygen species in c6 cells

Abstract

Central nervous system dysfunctions are among the most significant effects of exposure to ethanol and the glial cells that play an important role in maintaining neuronal function, are extremely involved with these effects. The actin cytoskeleton plays a crucial role in a wide variety of cellular functions, especially when there is some injury. Therefore the aim of the present study was to analyze the short-term effects of ethanol (50, 100 and 200 mM) on the cytoskeleton of C6 glioma cells. Here we report that acute ethanol exposure profoundly disrupts the actin cytoskeleton in C6 cells decreasing stress fiber formation and downregulating RhoA and vinculin immunocontent. In contrast, microtubule and GFAP networks were not altered. We further demonstrate that anti-oxidants prevent ethanol-induced actin alterations, suggesting that the actions of ethanol on the actin cytoskeleton are related with generation of reactive oxygen species (ROS) in these cells. Our results show that ethanol at concentrations described to be toxic to the central nervous system was able to target the cytoskeleton of C6 cells and this effect could be related with increased ROS generation. Therefore, we propose that the dynamic restructuring of the cytoskeleton of glial cells might contribute to the response to the injury provoked by binge-like ethanol exposure in brain.

Mechanical Stretch Inhibits Lipopolysaccharide-induced Keratinocyte-derived Chemokine and Tissue Factor Expression While Increasing Procoagulant Activity in Murine Lung Epithelial Cells

“…EtOH can modulate LPS-mediated cell signaling through actin reorganization (23,34,35). In a study by Dai and Pruett (23), exposure of the human macrophage cell line RAW264.7 to LPS in the presence of EtOH altered the distribution of TLR4 and its coreceptor, CD14, such that LPS-induced TNF-␣ was reduced.…”

Alcohol stress, membranes, and chaperones

“…Acute ethanol treatment resulted in disruption of the actin network and enhanced ROS formation in the C6 rat glioma cell line. The finding that different antioxidants can prevent ethanol-induced alterations in actin organization an indication that an elevated ROS level contributes to the cytoskeletal remodeling effect of ethanol (Loureiro et al 2011).…”

Dual action of chronic ethanol treatment on LPS-induced response in C6 glioma cells

“…Surprisingly, we found that regarding this parameter, C6 cells were not responsive to the effects of either EtOH alone or LPS alone, evaluated by DCF-DA measurement. Conversely, recently we found that C6 cells acutely treated with 50 to 200 mM EtOH developed increased ROS levels (Loureiro et al, 2011). Otherwise, it is interesting to note that EtOH plus LPS stimulation greatly increased oxidative stress generation, (Fig.…”

“…With successive passages (beyond passage 70) C6 glioma cell differentiates into a primarily astrocytic cell like population (Parker et al, 1980) and exhibits many properties of astrocytes, including expression of the astrocyte specific markers GFAP (Bissell et al, 1974) and S100B protein (Nardin et al, 2007). Moreover, we have recently reported the effects of acute treatment with EtOH in C6 glioma cell on actin cytoskeletal organization and we found that this remodeling could be related with oxidative stress (Loureiro et al, 2011).…”