Figure 3

CBD decreased reactive oxygen species in lesioned hemispheres as indicated by intensity of dihydroethidium staining (DHE, in red). (A) Representative confocal images displaying regional DHE staining indicative of reactive oxygen species (ROS) as a function of lesion condition and treatment. (B) Summary of mean Corrected Total Cell Fluorescence (CTCF) of DHE staining within each area using a circular area with a diameter of 0.5 mm (see methods for CTCF calculation details). In the vehicle group, microlesions significantly increased total fluorescence of DHE staining within HVC, RA and Area X. Significantly increased lesion-induced DHE staining was not observed in CBD treated groups. In all brain regions of lesioned hemispheres, DHE staining was significantly lower in CBD-treated groups compared to controls. (C) Comparing % control transformed measures of DHE staining within lesioned hemispheres of vehicle- and CBD-treated animals, CBD reduced intensities within HVC, RA, and Area X. CBD treatment significantly reduced lesion related changes in DHE expression in all regions as a percentage of their respective control hemisphere. Mean grey values were determined and corrected for background fluorescence using the average intensity outside of each song region studied. Comparisons were made and significance determined using Sidak’s correction for multiple comparisons following mixed-models ANOVA (n = 4 per group).