Fig. 2

Behavioral phenotypes in zebrafish ASD gene mutants

(A and B) Experimental setup and behavioral assays. Visual-startle: Larvae are exposed to five 1-s flashes of lights-off or -on stimuli at 29-s intervals at 5 dpf. Sleep-wake: Larvae are exposed to a 14 h:10 h white light:dark schedule at 5–7 dpf. Visual-startle and sleep-wake parameters are shown (see STAR Methods).

(C) Hierarchical clustering of mutant behavioral fingerprints. Each rectangle represents the signed -log₁₀-transformed p values from linear mixed models (LMM) comparing mutant and background-matched wild-type fish (red, increased in mutant; blue, decreased in mutant). Behavioral parameters from startle-off (light blue), startle-on (orange), and sleep-wake (purple) are shown. All mutant lines are shown except katnal2, which lacks consistently significant behavioral features (Figure S2A). Three mutant lines andtransheterozygotes (scn1lab^Δ44/Δ5) are shown for scn1lab. Behavioral features shown in individual graphs (D–G) are outlined in cyan. Mean and SD values for all groups and beta and p values for all behavioral parameters are shown in Table S2.

(D–G) Significant behavioral phenotypes in ASD gene mutants: increased responses to lights-on stimuli, daytime hypoactivity, and night-time hyperactivity in scn1lab^Δ44/Δ44 (D); decreased responses to lights-off stimuli and daytime hypoactivity in dyrk1aa^Δ77/Δ77dyrk1ab^Δ8/Δ8 (E); decreased daytime sleep bout length in chd8^Δ5/Δ5 (F); and daytime hyperactivity in tbr1a^Δ64/Δ64tbr1b^Δ10/Δ10 (G). Sleep-wake time series graphs in (D–G) show a rolling average of activity data points every 50 min. ^∗∗∗∗p < 0.0001, ^∗∗∗p < 0.001, ^∗∗p < 0.01 (one-way ANOVA).

(H) Correlation analysis across all 24 behavioral parameters identifies three sub-groups with related phenotypes: (i) scn1lab^Δ44/Δ44, chd8^Δ5/Δ5, cntnap2a^Δ121/Δ121cntnap2b^31ins/31ins; (ii) pogza^Δ23/Δ23pogzb^Δ20/Δ20, dyrk1aa^Δ77/Δ77dyrk1ab^Δ8/Δ8, cul3a^Δ7/+cul3b^Δ32/Δ32; and (iii) tbr1a^Δ64/Δ64tbr1b^Δ10/Δ10, kdm5ba^Δ17/Δ17kdm5bb^Δ14/Δ14, grin2ba^Δ25/Δ25grin2bb^Δ64/Δ64.