Fig. 4
β-glucocerebrosidase inactivation in oligodendrocytes causes demyelination in adult mice and affects oligodendrocyte differentiation in vitro. a, b Representative western blot (a) and relative densitometric quantification (b) of total brain protein extracts from Gba1f/f and Gba1f/f::cre mice of 6 months of age. MAG, PLP1 and MBP myelin protein levels were analyzed. β-actin (ACTB) was used as loading control (*, p < 0.05; unpaired two-tailed Student’s t-test; MAG, n = 14 mice; MBP, n = 15 mice for Gba1f/f and n = 14 mice for Gba1f/f::cre; PLP1, n = 15 mice for Gba1f/f and n = 13 mice for Gba1f/f::cre). Error bars indicate s.e.m. c, d Representative immunofluorescence images (c) and quantification (d) for MAG (top) and Fluoromyelin (bottom) in the striatum of 6-month-old Gba1f/f and Gba1f/f::cre mice. Nuclei were stained by Hoechst (**, p < 0.01; *, p < 0.05 unpaired two-tailed Student’s t test; n = 6 randomly selected fields from 2 mice per genotype). Error bars indicate s.e.m. Scale bar = 50 μm. e Representative electron microscopy images of striatal and optic nerve axons from 6-month-old Gba1f/f and Gba1f/f::cre mice. Scale bar = 1 μm. f, g G-ratio distribution of striatal and optic nerve myelinated axons (left panels), quantification of average g-ratio of myelinated axons (middle panels) and g-ratio distribution across axons divided into classes based on their diameter (right panels) (*, p < 0.05; **, p < 0.01; unpaired two-tailed Student’s t-test; striatum, n = 962 axon for Gba1f/f from 3 mice and n = 641 axons for Gba1f/f::cre from 3 mice; optic nerves, n = 459 axon for Gba1f/f from 3 mice and n = 624 axons for Gba1f/f::cre from 3 mice). Error bars indicate s.e.m. h, i Representative immunofluorescence for SOX10 in corpus callosum (CC) and striatum of 6-month-old Gba1f/f and Gba1f/f::cre mice (h) and relative quantification (i). Scale bar = 50 μm (n = 4 mice per group). Error bars indicate s.e.m. j, k Representative immunofluorescence images for MBP on differentiated primary oligodendrocytes (j) and relative quantification of MBP+ covered area (k). Scale bar = 50 μm (*, p < 0.05; unpaired two-tailed Student’s t-test; n = 3 oligodendrocyte cultures derived from a single pup). Error bars indicate s.e.m. l, m Representative immunofluorescence image for MAG on differentiated primary oligodendrocytes (l) and relative quantification of the percentage of mature MAG+ oligodendrocytes on total MAG+ cells, based on their ramified morphology (m). Scale bar = 100 μm. (*, p < 0.05; unpaired two-tailed Student’s t-test; n = 4 oligodendrocyte cultures derived from a single pup). Error bars indicate s.e.m