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Fig. 2  | Acta Neuropathologica Communications

Fig. 2 

From: Tau seeding in cases of multiple sclerosis

Fig. 2 

Anatomic distribution of tau seeding in an MS subject. The brain of an MS subject was preserved frozen, and then dissected to the indicated regions. Indicated regions were fixed for immunohistochemistry. For the tau seeding assay, unfixed tissue was homogenized to create total clarified lysate [10% (wt/vol)] followed by immunoprecipitation with MD3.1 to enrich for tau seeds. A Axial FLAIR MRI antemortem images showed extensive demyelination. B Periphery of an MS plaque was stained with Luxol fast blue-PAS-hematoxylin, showing preserved myelin in adjacent brain (left) and loss of myelin within the plaque (right). The plaque also contained abundant macrophages, and the interface (indicated with arrowheads) between plaque and adjacent brain contained many swollen axons. Scale bar = 1 mm. (C-J) Tau immunohistochemistry (AT8 and MD3.1) in plaque-adjacent brain regions. Scale bars = 50 µm. C, D temporal lobe, showing a collection of AT8-immunoreactive neuropil threads and MD3.1-immunoreactive tangle-like structures in 2 neurons; E, F parietal lobe, showing MD3.1-immunoreactive structures at the periphery of a plaque (consistent with swollen axons), but no AT8 immunoreactivity; G, H hippocampus, showing AT8-immunoreactive neuropil threads in entorhinal cortex, which are MD3.1-negative; and I, J substantia nigra, showing sparse AT8-immunoreactive neuropil threads, but no focal MD3.1 immunoreactivity. K Tau seeding in total clarified lysate from various regions of an MS brain. L Tau seeding in pellets after immunoprecipitation with MD3.1. Columns represent the mean % FRET positive cells from three technical replicates (dots). Statistical significance was determined by performing one-way ANOVA followed by Dunnett’s multiple comparisons testing of all samples vs. Lipofectamine treated negative controls, *** = p < 0.001, **** = p < 0.0001. Errors bars = S.D

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