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Fig. 5 Figure 5Glutamate Accumulation in akr1a1b-/- Mutants Damaged the Kidneys (A) Primary metabolites were measured in 96-hpf larvae by gas chromatography-mass spectrometry analysis and showed a significant increase for glutamate and alanine in ?+17 akr1a1b-/- larvae (n = 3 clutches with 50 larvae, mean ± SD). (B) Glutamate accumulated in adult ?+17/?-23 akr1a1b-/- zebrafish kidneys at 3 and 18 h postprandial (2 h postprandial: n = 7 in akr1a1b+/+, n = 8 in akr1a1b-/-; 3 h postprandial: n = 9 in akr1a1b+/+, n = 6 in akr1a1b-/-; 18 h postprandial: n = 10 in both groups, mean ± SD). (C) Enlarged glomerulus and shortened tubular neck length in 48-hpf wild-type zebrafish embryos treated with glutamate (n = 46 in control group; n = 50 in 0.1 mM group; n = 49 in 1 mM group, mean ± SD). (D) Representative pronephros images of glutamate treated 48hpf embryos. Glomeruli are encircled. (E) A significant increased loss of fluorescence in 1 mM glutamate-treated wild-type larvae both at 24 and 48 hpi (n = 26 in control group; n = 21 in 1 mM group, mean ± SD). ?p < 0.05, ??p < 0.01, ???p < 0.001, ????p < 0.0001; p value in (A, B, and E) was calculated by t test, p value in (C) was calculated by one-way ANOVA, Scale bar, 0.1 mm.