Figure 4

CBD alters inflammation-related cytokine densities. Daily treatments with 10 mg/kg were associated with modulation of IL-1Β, IL-6 and IL-10 protein expression 24 h after unilateral HVC lesions. Shown are representative immunofluorescent confocal images of antibody staining targeting IL-1Β, IL-6 and IL-10 within motor (HVC & RA) and learning-essential (Area X) regions. (Aa-Al), lesioned hemisphere images showing representative regional distribution of IL-6 in vehicle- vs. CBD-treated birds. (Ca–Cl), images demonstrating regional distribution of IL-1Β. (Ea-El), confocal images demonstrating regional distribution of IL-10. Representative images for the unlesioned hemisphere are presented in Supplemental Fig. S3. B, IL-6 fluorescence as percent change from unlesioned control hemisphere by song region and drug treatment. CBD significantly reduced IL-6 expression in HVC, RA, and Area X. D, IL-1B fluorescence as percent change from unlesioned control hemisphere by song region and drug treatment. CBD significantly decreased IL-1Β intensity in HVC, and RA. F, anti-inflammatory IL-10 fluorescence as percent change from unlesioned control hemisphere by song region and drug treatment. CBD significantly increased IL-10 intensity in RA and Area X. Differences were determined by mixed-models ANOVA followed by Sidak-corrected post-hoc comparisons. Image J software was used to analyze z-stack images projected at maximum intensity and threshold was applied consistently across all images for n = 5 per group.