Fig. 5

RFs visualized with Fluoro-Jade B (FJB) (a, b) and DAPI combined with immunohistochemical staining (c-d) in 1 year-old TG mice. a Subpial area rich in FJB+ RFs (bright green profiles). Note large size of RF aggregates in the vicinity of the pia (double headed arrows). Arrow indicates astrocyte without RFs visualized with FJB in cell body and processes. a1 Enlarged boxed area in (a) shows an astrocyte with many RFs in the form of small, bright puncta. b Variability of astrocytes in subpial area. In (b1) (enlarged boxed area in b), with DAPI counterstaining, which stains nuclei but not RFs, in combination with FJB. The astrocyte with many RFs (asterisk) is surrounded by astrocytes (arrows) without RFs. c Subpial area with a high level of astrogliosis. Double immunostaining for GFAP and CD44, counterstaining with DAPI. Confocal microscopy. c1 Enlarged boxed area in (c). Note: 1) small bright profiles (arrows in (c1)”, marked only some) in astrocyte bodies and processes corresponded to RFs stained with DAPI, 2) Top astrocyte (in c1) does not stain for CD44, but bottom left astrocyte does. Both contain many RFs. d) Aggregates of RFs in astrocyte processes near the pial surface. d1 –enlarged boxed area in (d). Large irregular profile of astrocyte process filled with many RFs that reveal minimal GFAP immunostaining of the central part. For animation of Z-stack of optical slices in image (d1) see Additional files 5 and 6: Movie 1 and 1a. Immunostaining for GFAP, counterstaining with DAPI. Black and white image show DAPI staining. Confocal microscopy. Scale bars: 60 μm in (a)-(c); 25 μm in (d)