Figure 5.

Chronic active MS lesions show Caspr re-distribution, increased NgR1 and axonal damage. For detailed methods please see the Supplementary Information. (A) Representative LFB-PAS stained images obtained from progressive MS patient brain tissues. From these, we selected periplaque (PP) and plaque (P) regions to further study Caspr distribution in these areas (scale bar = 500 μm). (B) In MS-PP, the co-localization of Caspr with juxtaparanodal Kv1.2 was observed. In MS-P, a significant elongation of Caspr+ segments and disrupted localization of Kv1.2 was observed (scale bar = 10 μm). (C) The ratio between the measured length of Caspr+ segments vs. their diameter was significantly increased in MS-PP and MS-P, compared with non-neurological disease control (NNDC) samples. (D) Immunostaining of SMI-32, Caspr, and DAPI in serial section showed a re-distribution of Caspr along the whole internode in SMI-32 (+) degenerative axons within the lesion border (white arrowheads indicate diffuse expression of Caspr along the internode; scale bar = 10 μm; one-way ANOVA with post hoc Tukey’s test, ****P < 0.0001, n = 4 for each patient samples). (E) Western blot for Caspr and (F) α-tubulin-loading control performed on brain white matter lysates of NNDC, Alzheimer’s disease (AD), frontotemporal dementia (FTD), Huntington’s disease (HD), and MS patients. (G) Densitometric quantification of full-length Caspr (FL-Caspr), (H) 64kDa degradation product of Caspr and (I) 45 kDa Caspr degradation product normalized by α-tubulin loading control (one-way ANOVA post hoc Tukey’s test, *P < 0.05, **P < 0.01, ***P < 0.001, n = 4 for each patient samples). (J) Immunoprecipitation of Caspr and probed with anti-PrP^C. Western immunoblot for PrP^C from 5% input of pre-immunoprecipitation sample shown on the bottom (K) Densitometric quantification of Caspr bound PrP^C and (L) total di-PrP^C. (M) Western blot for Reelin. (N) Densitometric quantification of 140 kDa Reelin.