FIGURE 6.

Ethanol decreases mitochondrial membrane potential via the activation of Panx1 and Cx43 hemichannels in astrocytes. (A–D) Representative photomicrographs of MitoRed CMXRos (red) and MitoGreen (green) fluorescence by astrocytes under control conditions (A, B) or following the treatment with 100 mM ethanol for 72 h (C,D). Insets: × 2.5 magnification of the indicated area of panels (B, D). (E) Averaged data of MitoRed CMXRos normalized to MitoGreen fluorescence by astrocytes under control conditions (white violin) or treated for several time periods with 100 mM ethanol alone (red violin). Also, it is shown the effect of the following pharmacological agents in astrocytes treated with 100 mM ethanol for 72 h: 100 µM ¹⁰panx1 (blue violin), 500 µM probenecid (Prob, green violin) or 100 µM gap19 (yellow violin). **p < 0.01, ***p < 0.001; ethanol compared to control conditions; ^& p < 0.05, ^&& p < 0.01; effect of pharmacological agents compared to ethanol treatment (one-way ANOVA followed by Tukey’s post hoc test). Each dot represents an independent culture experiment from a different litter of animals. The value reflects the mean of at least three coverslip replicates, with a minimum of 60 cells analyzed per coverslip. Calibration bar: 10 μm.