Fig 3

Zebrafish Brain TH levels increase with age and ethanol withdrawal.

Normalized TH levels in brain homogenates from zebrafish treated with 1.5% ethanol (black bars) or water (control, gray bars) plotted across caffeine dose (0 mg/L; 25 mg/L, 100 mg/L). (A) TH levels in brain tissue assessed immediately after exposure. Fish exposed at 140–149 dpf had increased TH levels relative to other exposure ages (p<0.0.006; asterisk). (B) TH levels were also compared across time of sacrifice—immediately after exposure, after the short post-exposure interval, or after the long post-exposure interval. Tissue collected immediately had significantly greater TH levels compared to tissue collected after the short interval or long interval (p<0.0037; asterisk). The boxplots show the minimum, first quartile, median, third quartile, and maximum values for each measure after outliers were removed. Exposure ages (dpf) are indicated at the top of the bar graphs. For brain TH levels, there were 4–6 independent replicates per treatment condition (each ethanol/caffeine dose) for a total of 35 subjects. ANOVA tables and multiple comparison results for this data can be found in S1 Table, panels (G1, G2) Brain TH levels across pre-exposure age; (H1, H2) Brain TH levels across time of sacrifice.