Figure 3

Immunolabeling of CA with the markers p62, GS and MMP2. Representative images of human brain hippocampus sections immunostained with p62 (a), GS (b) and MMP2 (c1 and c2). The three markers positively stained the CA (arrows). p62 also stained some dystrophic neurites (arrowheads in a), while MMP2 stained some thin filaments (arrowheads in c2). Hoechst (blue) was used for nuclear staining. d1–d2: double immunostaining with anti-ubiquitin (Ubi) (red) and p62 (green); e1–e2: double immunostaining with Ubi (red) and GS (green); f1–f2: double immunostaining with GS (green) and p62 (red). In some CA, p62 and GS are clearly visible in the peripheral region (d2, e2 and f2), while ubiquitin is concentrated in the central zone (d2 and e2). The different color channels from d2, e2 and f2 are shown in small images next to the corresponding ones. The histogram profiles of green and red intensities on the plotted lines illustrate the peripheral location of p62 and GS, and show that ubiquitin is concentrated in the central zone but is also present at the periphery. Double staining with GS and GFAP showed that GFAP staining is surrounding that of GS, which indicated that the astrocytic processes are surrounding CA (g1). This is clearly shown when CA is magnified (g2). When the histogram profiles are traced, it can be observed that the peaks of intensity of GFAP staining appear externally to those of GS staining. In some cases, CA become sliced in a tangential plane, and then the GS staining appear throughout the surface of the CA, and the astrocytic processes stained with GFAP are surrounding this surface (g3). Scale bars on d2, e2, f2, g2 and g3: 10 μm; other scale bars: 50 μm.