Fig. 6. Gaucher-like cells accumulate in gba1-/- and gba1-/-:asah1b-/- brains. A and B: WT, gba1-/-, and gba1-/-:asah1b-/ - brains were serially sectioned in the sagittal plane and H&E stained. Gaucher-like cells occupy the periventricular zone of the optic tectum in both gba1-/-I and gba1-/-:asah1b-/- brain, with higher magnifications of the boxed areas in (A) and (B). Scale bars: 200 μm and 50 μm for the low and higher magnifications, respectively. C: Transverse view of WT, gba1-/-, and gba1-/-:asah1b-/ - brains showing severe bilateral and diffuse accumulation of Gaucher-like cells in the periventricular gray zone of the optic tectum (HE). D: Immunohistochemistry for the zebrafish monocyte/macrophage lineage cells marker L-plastin. Immunoreactive macrophages were abundantly present in the ventricle and periventricular regions of gba1-/- and gba1-/-:asah1b-/- brains but not in the WT brain; counterstaining was performed using hematoxylin. Scale bars: 100 μm in C and D. E: mRNA expression of th1, th2, sncβ, sncγa, sncγb, and mbpa was determined using RT-qPCR analysis; WT (n = 7), gba1-/- (n = 9), asah1b-/- (n = 4), and gba1-/-:asah1b-/- (n = 7). Data of RT-qPCR is normalized using two housekeeping genes (ef1α and rpl13), analyzed by One-Way Anova with Tukey’s multiple comparisons test or a Brown-Forsythe and Welch Anova with Dunnett’s multiple comparisons test for gpnmb and chia.6 and depicted as scattered dot plot ± SEM. 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. Ns = not significant, ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗∗p < 0.0001.
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