Fig 5

Zebrafish Brain GAD65/67 levels vary with age.

Normalized brain GAD65/67 levels of zebrafish exposed to 1.5% ethanol (black bars) or water (gray bars) ± caffeine (0 mg/L; 25 mg/L, 100 mg/L). (A) There was a significant effect of pre-exposure age on brain GAD levels. GAD65/67 levels in brain tissue collected immediately after exposure showed a trend of increased levels when fish were exposed earlier, from 50–59 dpf, compared to later ages (60–149 dpf). (B) GAD65/67 levels assessed across the post-exposure time points showed a trend of greater GAD65/67 levels in subjects exposed after a long interval compared to those sacrificed immediately. 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 across the top of the bar graphs. For brain GAD 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 (L1, L2) Brain GAD levels across pre-exposure age; (M) Brain GAD levels across time of sacrifice.