Integrated patient induced pluripotent stem cell models and Gaa-/- mice reveal central nervous system neural pathology in infantile-onset Pompe disease

Figure 4 Progressive neurodegeneration in Gaa^-/- mice and patient-derived neural cultures. A: Representative GFAP immunofluorescence images of the cerebral cortex from 3- and 6-month-old wild-type (WT) and Gaa^-/- mice. Scale bar = 100 μm; B: Representative GFAP immunofluorescence images of the spinal cord from 3- and 6-month-old WT and Gaa^-/- mice. Scale bar = 100 μm; C: Quantification of GFAP fluorescence intensity in the cerebral cortex shown in A (n = 4 mice per group; data are presented as mean ± SEM; ^aP < 0.05, WT 3 months vs Gaa^-/- 3 months; ^aP < 0.05, WT 6 months vs Gaa^-/- 6 months; ^aP < 0.05, Gaa^-/- 3 months vs Gaa^-/- 6 months; one-way ANOVA); D: Quantification of GFAP fluorescence intensity in the spinal cord shown in B (n = 4 mice per group; data are presented as mean ± SEM; ^bP < 0.01, WT 3 months vs Gaa^-/- 3 months; ^dP < 0.0001, WT 6 months vs Gaa^-/- 6 months; ^aP < 0.05, Gaa^-/- 3 months vs Gaa^-/- 6 months; one-way ANOVA); E: Representative NeuN immunostaining of the cerebral cortex from 3- and 6-month-old WT and Gaa^-/- mice. Scale bar = 100 μm; F: Representative NeuN immunostaining of the spinal cord from 3- and 6-month-old WT and Gaa^-/- mice. Scale bar = 100 μm; G: Quantification of the NeuN+ area fraction in the cerebral cortex shown in E (n = 3 mice per group; data are presented as mean ± SEM; ^aP < 0.05, WT 6 months vs Gaa^-/- 6 months; one-way ANOVA); H: Quantification of the NeuN+ area fraction in the spinal cord shown in F (n = 3 mice per group; data are presented as mean ± SEM; ^aP < 0.05, WT 6 months vs Gaa^-/- 6 months; ^aP < 0.05, Gaa^-/- 3 months vs Gaa^-/- 6 months; one-way ANOVA); I: Immunofluorescence images of patient - derived induced pluripotent stem cell neurons cultured for 90 days, showing increased cleaved caspase-3 (red) positivity in TUBB3+ neurons (green) compared to controls. Nuclei counterstained with DAPI (blue). Scale bar = 20 μm; J: Quantification of cleaved caspase-3 positivity in TUBB3+ neurons shown in I (n = 3 independent differentiation; data are presented as mean ± SEM; ^aP < 0.05; unpaired two-tailed Student’s t-test). WT: Wild-type; GAA: Acid α-glucosidase; IOPD: Infantile-onset Pompe disease.