Progression of metabolic abnormalities in 10 dpf old alpl-/- zebrafish larvae. (a) Kaplan–Meier survival curves of untreated and 100 µM PN-treated WT and alpl-/- zebrafish until 10 dpf. Data are from n = 18–42 zebrafish per genotype and treatment condition. (b) Concentrations of pyridoxine (PN), PLP, pyridoxal (PL) and its degradation product 4-pyridoxic acid (4-PA) in untreated and 100 µM PN-treated WT and alpl-/- larvae. Data are means from n = 4–10 larvae per genotype and treatment condition ± SD. (c) Distance swum during burst activity (lardist, movement speed > 30 mm/s) and during small activity (smldist, movement speed < 30 mm/s) during 1 h measurement using Zebrabox at 28 °C in the dark. Data are means from n = 9–19 larvae per genotype and treatment condition ± SD. (d) Concentrations of GABA in untreated and 100 µM PN-treated WT and alpl-/- larvae. Data are means from n = 5–7 larvae per genotype and treatment condition ± SD. (e) Important features in metabolome of alpl-/- larvae selected by volcano plot with fold change (FC) threshold equal to 2 (x-axis) and t-tests threshold of p < 0.05 (y-axis). (f) Accumulation of L-Dopa and vanillactic acid, and decreased level of dopamine, dopamine sulfate (dopamine 4-sulfate and dopamine 3-O-sulfate) and epinephrine in alpl-/- larvae suggest impaired activity of aromatic L-amino acid decarboxylase. (g) Accumulation of vitamin A (retinol) and decreased level of retinal in alpl-/- larvae. (h) Accumulation of N-methylethanolaminium phosphate in alpl-/- larvae. Data are means from n = 5–7 larvae per genotype ± SD (panels E-H). **** p < 0.0001, *** p < 0.001, ** p < 0.01, and * p < 0.05.
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