adgrl3.1^−/− shows attention deficits, impulsivity, and increased risk-taking.

A Flow-chart summarizing the 5-CSRTT process. During the 5-CSRTT, fish were required to swim toward one of five spatially distinct LEDs when illuminated. Approaches to the illuminated light were ‘correct’ and the proportion of correct trials was a measure of attention. Prior to illumination, there was a variable-time (mean 5-s) inter-trial interval, and responses during this interval were punished with subsequent non-reinforcement. Responses during this inter-trial interval (anticipatory or premature responses) were used as a measure of impulse control. B No significant effects were found after two-way RM ANOVA for the acquisition during the Stage IV of the 5-CSRTT (Day*Group effect—F(8, 136) = 0.50; p = 0.95; Group effect—F(1, 17) = 0.69; p = 0.41; Day effect—F(4, 247) = 0.45; p = 0.77). C One-way ANOVA yielded a significant effect for accuracy (F(2,25) = 5.80; p** = 0.0085), D anticipatory responses (F(2,25) = 14.17; p**** < 0.0001) with no effects for (E) omissions (F(2,25) = 1.80; p = 0.18). Tukey’s post-hoc analysis was used to characterize significant differences (p** < 0.005 and p**** < 0.0001; n = 9–10). F Risk-taking behavior is defined as time spent close to the novel object. A significant ANOVA effect was observed for time spent close to the object (F(2,32) = 8.35; p** = 0.0012) where adgrl3.1^−/− spent more time close to the object (p* = 0.0148), an effect that was 750 significantly decreased by atomoxetine in adgrl3.1^−/− (p** = 0.015; n = 12–13). The data is represented as mean ± S.E.M.

adgrl3.1^−/− zebrafish show increased locomotion after 3 days of habituation.

A Significant two-way RM ANOVA effect for habituation (F(1.995, 135.7) = 3.84; p* = 0.024) was observed for distance traveled, with no significant effects for interaction between factors (genotype*habituation; F (2, 136) = 1.66; p = 0.19) nor genotype (F (1, 68) = 1.40; p = 0.24). For immobility, a significant effect of genotype*habituation (F (2, 136) = 3.66; p* = 0.03), habituation (F (1.998, 81.51) = 5.92; p* = 0.013), and genotype (F (1, 68) = 10.23; p** = 0.002) was found. Post-hoc analyses showed that adgrl3.1^–/– has increased immobility during the first day of habituation compared to WT (p* = 0.018). adgrl3.1^−/− also showed a habituation to the novel environment by showing a decrease in their immobility during the second day (p* = 0.025) and third day (p* = 0.03) compared to the first day of habituation. B Representative tracking of a WT vs. adgrl3.1^−/− vs. adgrl3.1^−/− + ATO animal during the test day. C A significant ANOVA effect was found for distance traveled (F(2,48) = ^5.44; p** = 0.007) during the test day. Briefly, distance traveled was increased for adgrl3.1^−/− compared to WT animals (p** = 0.005) with no effect for adgrl3.1^−/− compared to adgrl3.1^−/− + ATO (p = 0.14). No effect for immobility was observed (p = 0.54). The data is represented as mean ± S.E.M.

RNA-seq summary data.

A Principal component analysis plot of the top 200 most variable genes after differential expression analysis. n = 4 brains/group. B Venn diagram showing the overlap of differentially expressed genes (DEGs) between WT zebrafish, adgrl3.1^−/− and adgrl3.1^−/− treated with atomoxetine. C Heatmap of DEGs (lowest adjusted p value (p_adj) < 0.05 and LFC > |2 | ) between WT and adgrl3.1^−/−. D Volcano plot displaying DEGs between WT and adgrl3.1^−/−. DEGs with p_adj are highlighted. n = 4 per group. E Heatmap of DEGs (p_adj < 0.05 and LFC > |2 | ) between WT and adgrl3.1^−/− treated with atomoxetine. F Volcano plot displaying DEGs between WT and adgrl3.1^−/− treated with atomoxetine. DEGs with p_adj are highlighted. n = 4 per group. G Heatmap of DEGs (p_adj < 0.05 and LFC > |2 | ) between adgrl3.1^−/− and adgrl3.1^−/− treated with atomoxetine. H Volcano plot displaying DEGs between adgrl3.1^−/− and adgrl3.1^−/− treated with atomoxetine. DEGs with p_adj are highlighted. n = 4 per group.