Behavioral phenotypic screening of candidate ASMs in WT and scn1lab larvae. (A) Representative baseline swim velocities (mm/s) over time for 5 dpf WT and scn1lab mutant larvae. Threshold for behavioral seizure events indicated by dotted line (28 mm/s). Scn1lab larvae (N = 60 across 3 replicates) have significantly (P = 0.014) higher maximum velocities on average compared to WTs (N = 70 across 3 replicates), all larvae are represented as individual data points. (B) Timeline for behavioral tracking acquisition (top). Heatmap of the percent change in average swim velocity from baseline after treatment with candidate ASMs at three different concentrations. Significant changes from vehicle control are indicated by stars with an N = 30 or 40 per condition across minimum three trials for WT and scn1lab larvae. (C) Behavioral tracking plots for WT larvae showing baseline activity followed by ASM treatment for drugs that caused a significant change in swimming velocity compared to DMSO controls. The plot below highlights the normalized velocity in percent, after ASM treatment for each larvae recorded. AA43279 significantly increased swim velocity in WT larvae at 100 µM (N = 40 across three replicates, P < 0.0001). (D) Behavioral tracking plots for scn1lab larvae showing baseline activity followed by ASM treatment for drugs that caused a significant change in swimming velocity compared to DMSO controls. Donepezil increased swim velocities for scn1lab larvae (N = 40 across 4 replicates, P < 0.01). 1-EBIO, chlorzoxazone and lisuride significantly reduced swim velocities (N = 30–40 across 3–4 replicates, P < 0.05–0.0001). The plot below highlights the normalized velocity in percent, after ASM treatment for each larvae recorded. An unpaired t-test or ANOVA was performed for statistical analysis, P < 0.05 = *, P < 0.005 = **, P < 0.0005 = ***, P < 0.0001 = ****.
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