Early seizure onset and death following Lysine (Lys) treatment. Survival of aldh7a1−/− and wild-type (WT) siblings after 48 hr of exposure with Lysine 20 mM (A) started at days 6, 7, or 8 days postfertilization (dpf). Survival of aldh7a1−/− and WT siblings after 48 hr of treatment with Lys with or without daily Pyridoxine 10 mM (Pyr) treatments (B). Survival experiments were terminated at 20 dpf. Video analysis of 9-dpf larvae after 24 hr of treatment with Lys 20 mM, Pyr 10 mM, or both, showing significant increase in duration (C) and distance moved (D) in high-speed movements after light stimulus. Representative movement traces obtained from the Zebralab software at 9 dpf (24 hr after Lys treatment) showing the presence of larvae with hyperactivity and high-speed movements (red) (E). Blinded analysis of the same videos analyzed in (C–E) showing classification of each fish by seizure scores (S0, S1, S2, and S3), n = 8 larvae per group, except for mutant + Lys 20 mM where two fish died during the first 24 hr of treatment (F). Asterisks on top of the graph indicate statistical significance according to one-way ANOVA test [P = 0.0081 in (C) and P = 0.0060 in (D)] and asterisks comparing each pair of samples reflect Tukey’s post hoc pairwise tests (* P < 0.05 and ** P < 0.01). Error bars represent ± SD.
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