Fig. 1

Cuprizone feeding induces demyelinated lesions with pronounced gliosis in the cerebellar nuclei with sparing of the cerebellar cortex. a. Cuprizone was administered for 5 consecutive weeks, followed by histological or electrophysiological analysis. b Schematic representation of the analyzed regions of the murine cerebellum, namely the cerebellar nuclei (CN) and the cerebellar cortex, consisting of the subcortical lobar white matter (WM), the granule cell layer (GCL) and the molecular layer (ML). Analysis of the cerebellar cortex was performed in the area of the vermis (v). c Representative overview micrographs depict the cuprizone-induced demyelinated lesion in the CN and the fully myelinated cortical region in LFB-PAS staining (left). Abundant MAC3⁺ activated microglial cells are detectable within the lesion. In the subcortical lobar WM and GCL close to the lesion, some activated microglial cells (red arrows) can be observed (middle image, MAC3 IHC). Astrogliosis is apparent in the lesion, to some extent reaching into the myelinated subcortical lobar WM and GCL close to the lesion (right image, GFAP-IHC). d Quantification of MAC3⁺ cells in different cerebellar regions, in and outside of the demyelinated lesion, comparing cuprizone-fed vs. naive mice. Higher magnification images (right upper panel) show the cerebellar cortex with only single MAC3⁺ cells (red arrow) in contrast to a massive microglial activation within the lesion (lower right panel). Each point represents a measurement from an individual animal (naive n = 4; CUP5W n = 6). e Quantification of GFAP⁺ area in different cerebellar regions of cuprizone-fed vs. naive mice. High magnification images (right panel) indicate GFAP⁺ Bergmann glia cells in naive (top) vs. cuprizone-fed mice (bottom) (GFAP-IHC). Each point represents a measurement from an individual animal (naive n = 7; wt CUP5W n = 8). Missing values in d (CN) correspond to insufficient cerebellar nuclei areas in two CUP5W samples. Whiskers represent mean ± SEM. P values were obtained after two-way ANOVA followed by Sidak’s multiple comparisons test. Asterisks represent significant p-values (*p < 0.05, **p < 0.01, ***p < 0.001, **** p < 0.0001)