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Fig. 5

Fig. 5. Biochemical abnormalities in adult zebrafish brain. A: Lipid levels were determined of dissected brains in pmol/mg tissue. GlcSph, GalCer, and GlcChol were separated from their respective galactosylated lipid by HILIC chromatography. Data is depicted as mean ± SEM; WT (n = 9–10), gba1-/- (n = 9), asah1b-/- (n = 8), and gba1-/-:asah1b-/- (n = 11). Data is analyzed by One-Way Anova with Tukey’s multiple comparison test. B: mRNA levels of asah1a, asah1b, gpnmb, chia.6, il-1β, tnfβ, apoEb, catD, c1qA, c3.1, c5aR, and c5 was determined using RT-qPCR analysis; WT (n = 9–11), gba1-/- (n = 8–11), asah1b-/- (n = 5), and gba1-/-:asah1b-/- (n = 9–10). Data is analyzed by One-way Anova with Tukey’s multiple comparison test. C: Representative Western blot of p62, LC3-I, LC3-II, β-catenin protein levels in WT, gba1-/-, asah1b-/-, and gba1-/-:asah1b-/- zebrafish brains, with β-actin as protein loading control. D: Quantitative analysis of LC3-II/LC-I levels and p62 protein levels. WT (n = 5), gba1-/- (n = 5), asah1b-/- (n = 3), and gba1-/-:asah1b-/- (n = 6). Statistical analysis is depicted of WT versus gba1-/-, asah1b-/-, or asah1b-/-:gba1-/-, only when a significant difference is apparent or of gba1-/- versus asah1b:gba1-/- when significant difference is of interest. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗∗ p < 0.0001. GalCer, galactosylceramide; GlcSph, glucosylsphingosine; GlcChol, glucosylated cholesterol.

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