Fig. 1

Cell viability after ethanol exposure is sex dimorphic in ED10.5 and this dimorphism is eliminated by inhibition of ALDH. (a) This cellular representation of alcohol metabolism shows the canonical alcohol metabolism pathway: ethanol enters the cell by diffusion and is metabolized by alcohol dehydrogenase in the cytoplasm to acetaldehyde. Aldehyde dehydrogenase (ALDH) further metabolizes the acetaldehyde to acetate, which is later used by the cell. The non-canonical alcohol metabolism pathway is similar to the canonical pathway, but the initial metabolic step is performed by Cyp2e1 on microsomes and results in generation of reactive oxygen species (ROS). Alcohol dehydrogenase (ADH) is the major enzyme pathway for oxidizing ethanol to acetaldehyde. (b) Cell death measured by trypan blue exclusion. In this and other figures, blue bars represent male cells and red bars, female cells. Cells from ED10.5 whole embryo in culture were exposed to 400 μM ethanol for 24 h. Data represent percentage total cell death for all conditions. These experiments were done in triplicate. Higher female cell sensitivity to ethanol is seen in cells. Asterisks (*) indicate p < 0.05 between male and female. The error bars represent S.D. (c) Measure of mitochondrial membrane integrity by WST-1 after exposure to 400 μM EtOH. Cells were seeded and incubated in 96-well dishes for the final hour of a 24-h exposure to EtOH with the Wst-1 compound and read at 450 nm with background subtracted at 690 nm. Male cells have higher WST-1 activity in the presence of EtOH for ED10.5 whole embryo cells. These experiments were done in triplicate, with asterisks (*) representing p < 0.05. (d) Confirmation of the WST-1 assay with MTT assay. Cell mitochondrial integrity after exposure to 400 μM EtOH for 24 h. Cells were supplemented with MTT stock for 2 h, and then the dye was solubilized and supernatants were read at 540 nm. Female cells are more sensitive than male cells to EtOH. These experiments were done in triplicate, with asterisks (*) representing p < 0.05. (E) Cell death measured by trypan blue exclusion. Cultured cells exposed to 400 μM ethanol, DSF or a combination. Higher female cell sensitivity to ethanol is seen in cells from ED10.5 whole embryo, but not in cells co-treated with DSF. Data represent percentage total cell death for each condition. These experiments were done in triplicate. Asterisks (*) represent P < 0.05