Adult bcas3 KO zebrafish showed increased anxiety, aggression and abnormal social. (A-C), Analysis of the behavior of 4 mpf WT and bcas3−/− zebrafish in novel tank test. (A), Representative trajectory and heatmap image. Quantitative analysis of distance (B) and time (C) spent in bottom, middle, and top zones. (n = 17, per group). Bcas3 KO zebrafish exhibited reduced distance and movement time spent in the top zone compared to WT controls, indicating increased anxiety-like behavior. The tank was divided horizontally into three equal sections: the top zone, the middle zone, and the bottom zone. (D-F), Analysis of 4 mpf WT and bcas3−/− zebrafish behavior in mirror biting test. (D), Representative trajectory and heatmap image. Quantitative analysis of average velocity (E) and active time spent in contact zone (the distance from the mirror < 2 cm), approach zone (2 cm ≤ the distance from the mirror ≤ 6 cm), and far zone (the distance from the mirror > 6 cm) (F) in WT and bcas3−/− zebrafish. ①, contact. ②, approach. ③, far. (n = 17 per group). Bcas3 KO zebrafish spent more time in the contact zone, indicating heightened aggressive behavior. (G-H), Analysis of 4 mpf WT and bcas3−/− zebrafish behavior in shoaling behavior test. Representative image (G) and average inter-fish distance (H) was significantly increased in bcas3−/− zebrafish, indicating reduced social cohesion. (n = 30, per group). (I-L), Analysis of 4 mpf WT and bcas3−/− zebrafish behavior in social test. (J), Representative trajectory and heatmap image. Quantitative analysis of total movement distance (I), average velocity (K) and time spent in left and right regions (L) in WT and bcas3−/− zebrafish. (n = 11, per group). Data are shown as mean ± SEM. Unpaired Student’s t-test was used to analyze movement distance, average velocity and active time spent in different zones. Significance levels are denoted as follows: *P < 0.05, **P < 0.01, ***P < 0.001. ns, not significant
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