Effect of saffron on induced-TBI zebrafish behavior in novel tank diving test on days 3, 10 and 21 post-lesion. (A) Time spent at top of the tank (s) in CTL-saffron (n = 14), TBI-saffron (n = 14), and TBI + saffron (n = 6) at 3-, 10- and 21-days post-injury. (B) Latency to enter the top (s) in CTL-saffron (n = 14), TBI-saffron (n = 14), and TBI + saffron (n = 6) at 3-, 10- and 21-days post-injury. Statistical analysis was performed with two-way ANOVA. Statistically significant differences between groups are indicated as * p < 0.05, ** p < 0.005, *** p < 0.001. All data represent the mean ± S.E.M.

Effect of saffron on induced-TBI zebrafish behavior in the light/dark test on days 3, 10 and 21 post-lesion. Time spent in the light area (s) in CTL-saffron (n = 14), TBI-saffron (n = 14), and TBI + saffron (n = 6) at 3-, 10- and 21-days post-injury. No statistically significant differences were noted using a two-way ANOVA analysis. All data represent the mean ± S.E.M.

Effect of saffron on induced-TBI zebrafish behavior in the predator test on days 3, 10 and 21 post-lesion. Time spent near the predator comparisons between CTL-saffron (n = 14), TBI-saffron (n = 14), and TBI + saffron (n = 6) at 3-, 10- and 21-days post-injury. Statistical analysis was performed with two-way ANOVA. Statistically significant differences are indicated as * p < 0.05, ** p < 0.005, *** p < 0.0001. All data represent the mean ± S.E.M.

Active avoidance test on zebrafish 12 days post-injury to evaluate the effect of saffron on learning, short-term memory (STM) and long-term memory (LTM). (A) Experimental set-up used for active avoidance test. (B) Comparison of the number of avoidance (per 6 trials) in CTL-saffron (n = 9), TBI-saffron (n = 7), and TBI + saffron (n = 6) at 12 days post-injury during training session S1, S2 and S3. Statistical analysis was performed with two-way ANOVA. Statistically significant differences are indicated as a (in S1: CTL-saffron vs TBI-saffron; p < 0.05); b (in S1: TBI + saffron vs TBI-saffron; p < 0.005); c (in S1: CTL-saffron vs TBI-saffron; p ≤ 0.0001 and in S2: CTL-saffron and TBI + saffron vs TBI-saffron; p ≤ 0.0001). Data represent the mean ± S.E.M. (C) Comparison of the number of avoidance (per 6 trials) in CTL-saffron (n = 9), TBI-saffron (n = 7), and TBI + saffron (n = 6) at 12 days post-injury during training sessions (average number during S1, S2 and S3 taken together), STM and LTM. Statistical analysis was performed with two-way ANOVA. Statistically significant differences are indicated as ** p < 0.005, *** p < 0.0001. Data represent the mean ± S.E.M. (D) Comparison of the number of trials (per session) that was necessary for zebrafish to effectuate the first successful avoidance in CTL-saffron (n = 9), TBI-saffron (n = 7), and TBI + saffron (n = 6) at 12 days post-injury during training sessions (average number during S1, S2 and S3 taken together; upper graph), STM (middle graph) and LTM (lower graph). Statistical analysis was performed with one-way ANOVA. Statistically significant differences are indicated as ** p < 0.005, *** p < 0.0001. Data represent the median ± S.E.M.

(A) Dorsal and (B) lateral view of an adult zebrafish brain illustrating the injury site (black arrow) in the right dorsal telencephalon. Tel, Telencephalon. Scale bar: 1 mm.