The use of CIE as a more accurate model of studying the effects of high alcohol exposure in vitro

Previously, exposure of cultured cells to high concentrations of ethanol (EtOH) has included complete removal of the EtOH-containing media after a set number of hours. Chronic Intermittent Exposure (CIE) is a more physiologically actuate way of representing the way alcohol and the human body interact. CIE involves removing 50% of the alcohol from cells, without leaving the cells dry, at two hour intervals throughout the day to mimic the body’s rate of alcohol breakdown. During our experiment, samples were taken at three separate time points; cover slips were stained and mounted to slides. Mitotracker was used to stain the mitochondria of astrocytes and phalloidin targeted the actin cytoskeleton of astrocytes. The slides will be examined with a confocal microscope and images will be taken. The images will be fluoresced and analyzed with Metamorph software to assess alterations in intracellular mitochondrial distribution and actin depolymerization due to CIE. An increase in actin depolymerization and/or perinuclear clustering of intracellular mitochondria is both indicators of cell death. It remains to be seen if CIE results in astrocytic death or if it is a viable alternative to study cultured cells exposed to high concentration of EtOH similar to those observed with a human pattern of binge drinking.