To address the possibility that blunted ethanol sensitivity in Drosophila Clic mutants could be secondary to altered ethanol uptake or metabolism, we determined internal ethanol concentrations in flies exposed to ethanol in eRING assays (Bhandari et al., 2009). Duration of ethanol exposure had a significant effect on internal ethanol concentrations as expected (individual two-way ANOVAs; Figure 4A, F(2,12)=115.69, p<0.0001; Figure 4B, F(2,12)=84.39, p<0.0001; n=3), but there was no effect of Clic genotype on internal ethanol concentrations (Figure 4A, F(1,12)=0.417, p=0.529; Figure 4B, F(1,12)=0.283, p=0.603) or an interaction between genotype and duration of ethanol exposure (Figure 4A, F(2,12)=0.340, p=0.569; Figure 4B, F(2,12)=0.888, p=0.362). Clic mutants, therefore, have a blunted level of response to acute ethanol exposure that is not related to a significant change in tissue concentrations of the drug.
