Association of severe demyelination with extensive microgliosis in the brain stem. (A-C) Triple staining of microglia (F4/80, green), damaged axons (SMI32, red) and myelin (MBP, blue). Already at the age of three and six weeks demyelinated and degenerating axons are observed in a subset of Nestin-Pex5
−/− mice (B-C), in contrast with the control littermates (A). Microglial cells are observed in association with these demyelinated axons (C, arrow). (D-F) Higher magnification of the double labeling of myelin (MBP, red) and microglia (F4/80, green) in the brain stem of six- to nine-week-old Nestin-Pex5
−/− mice. The observed microglial cells have a swollen appearance and several of them contain MBP-positive myelin debris (D-F, arrows). (G-I) The anti-GFAP antibody was used to analyze astrogliosis, which is mild at three weeks (H), but strongly pronounced at 12 weeks (I), compared with the control mice (G). All panels of each row were stained with the same antibodies. Scale bars: A-D and G-I: 100 μm; E-F: 50 μm. GFAP, glial fibrillary acid protein; MBP, myelin basic protein.