An In Vitro Model to Study Chronic Intermittent Exposure that is similar to Human Binge Drinking

These experiments were designed to replicate binge drinking in humans, a pattern of high alcohol consumption in a short period of time. To date, In vitro studies utilizing this pattern of high alcohol exposure do not exactly replicate human alcohol consumption in that the alcohol is completely removed after 18-20 hours and not slowly metabolized over time. To best replicate the human alcohol metabolic process in primary cultured cerebellar granule cells (CGC’s), we administered 100 mM EtOH at 1600 H. At two-hour intervals starting at 800 H the next day we replaced 50% volume of feeding media (FM) with conditioned (FM) containing no alcohol. This was done for four days followed by a four-day recovery period. At 800 H the day after each ethanol administration, mean alcohol values as determined by gas chromatography, were 44.4 + 5.3 mM. Ethanol vales were zero by the end of the day just prior to the next alcohol administration. To measure neuronal viability, the trypan blue exclusion method will be used to measure the percentage of dead cells attributed to alcohol exposure. Visually, the cells all seemed to remain healthy and alive throughout the entire experiment. Are initial conclusion is that this in vitro alcohol exposure paradigm is a true chronic intermittent exposure (CIE) paradigm similar to that observed during human binge drinking.