Figure 4

Nuclear TPPP staining is reduced in MSA oligodendroglia. a-c: Klűver-Barrera staining (a) and an immunohistochemistry for TPPP (TPPP-C-psB) in normal control pons (Case 19). Note that myelin structures mostly show TPPP-immunoreaction, though this correlation is not always true as in the medial lemniscus and superior cerebellar peduncle. A higher magnification of (b) show TPPP-immunoreactivity in neuropil and in nuclei (c). d-g: In MSA-pons (Case 8), loss of myelin structure in Klűver-Barrera staining (d) is also observed in an immunohistochemistry for TPPP (TPPP-C-psB) (e). Where the TPPP immunoreactivity is very low (f), TPPP-positive cells are very few, suggesting oligodendroglial cell loss. In an area with relatively stronger TPPP-immunoreactivity (g), TPPP-positive oligodendroglia are remaining. Note that MSA-oligodendroglia (f & g) tend to lack nuclear-TPPP immunoreactivity compared to control (c). h: A three-dimensional image of double-labeling immunofluorescence of an MSA-cerebellum (Case 1) using TPPP-N-mab#2A8G3 and the anti-lamin B1 antibody. Note that nuclear TPPP is absent and the TPPP-immunoreactive cytoplasm is widened in this cell. i: Double-labeling immunofluorescence demonstrating the localization of TPPP (TPPP-N-mab#2A8G3) and TDP43, or TPPP and OLIG2 in the MSA-cerebellum (Case 1). TDP43 and OLIG2 are retained in the nucleus, whereas TPPP is absent. j: Classification of oligodendroglia into six types based on the localization of TPPP (TPPP-N-mab#2A8G3) and pα-syn. Hoechst staining is omitted from this figure. k: Box and whisker plot demonstrating the frequency of oligodendroglia with TPPP immunoreactivity in the nucleus. Compared with the normal control, nuclear TPPP is significantly reduced in pα-syn-negative MSA-oligodendroglia (type 1/types 1+2). The reduction of TPPP in the nucleus was more pronounced in MSA-oligodendroglia with GCIs (types 3+5/types 3+4+5+6). Nuclear TPPP was also reduced in oligodendroglia containing TPPP-negative GCIs (type 5/types 5+6) than in those containing TPPP-positive GCIs (type 3/types 3+4). *p<0.05, (k). Scale bar=5 mm (a, b, d, e), 20 µm (c, f, g), 10 µm (h, j), 5 µm (i).