Behavioral (A, D, G) and electrophysiological (B, C, E, F, H, I) antiepileptic activity of combination treatment (colored in blue) of valproate (VPA) with stiripentol (STP) (A–C), (±) fenfluramine [(±)-FFA] (D–F), and cannabidiol (CBD) (G–I) in the zebrafish scn1Lab−/− mutant model. (A, D, G) Locomotor activity of larvae pre-exposed to antiepileptic drugs (AEDs) for 22 h. Data were assessed over the total tracking period of 10 min and expressed as cm/100 s. Results were pooled from 3–4 independent experiments, with 120 larvae for each of the VHC-treated groups, and 20–30 larvae for each AED-treated group. (B, C, E, F, H, I) Noninvasive local field potential (LFP) recordings from the optic tectum of larvae pre-exposed to antiepileptic drugs (AEDs) for 22 h. Epileptiform discharges are quantified by the cumulative duration (mean ± SEM) (B, E, H) and frequency (mean ± SEM) (C, F, I) of events per 10-min recording. With 23 larvae for the VHC-treated group, 12–14 larvae for each AED-treated group. Statistical analysis: one-way ANOVA with Dunnett’s multiple comparison test (locomotor assay), Kruskal–Wallis testing with Dunn’s multiple comparisons (LFP measurements). Significance levels: *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. WT wild type, VHC vehicle
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