Figure 5
11A1 immunoreactivity in protoplasmic astrocytes. (A and B) Representative images were obtained under 63x objective lens. Protoplasmic astrocytes of human inferior parietal cortex, layer V, immunostained with 11A1 and ALDH1L1 antibodies and 11A1, GFAP and Cathepsin D antibodies. The rows of figures correspond to three age groups: early 30 s, middle 40 s, and early 60 s. (A) Left panels show the 11A1 signal (green), middle panels correspond to the ALDH1L1 signal (red) and right panels show the merged images. Scale bar 5 µm. (B) Left panels show the 11A1 signal, middle panels correspond to the Cathepsin D signal (magenta) and right panels show the merged images. The white contours in the merged images indicate the cell shape identified by GFAP. Separate localization of the 11A1 immunoreactivity and Cathepsin D signals in the astrocytes is demonstrated. Scale bar 5 µm. (C) Verification of 11A1 antibody’s specificity against Aβ toxic conformer in protoplasmic astrocytes of the three age groups. The 11A1 antibody was pre-incubated with (bottom panels) or without (top panels) its immunogen, E22P-Aβ9-35ox peptide, prior to incubation of tissue sections with primary antibody. The pre-incubation with immunogen abrogates the 11A1 signal (green) in and around ALDH1L1 signal (magenta). Images were obtained under 20x objective lens. Scale bar 20 µm. (D) Percentage of protoplasmic astrocytes labeled with 11A1 of the astrocytes in measured area across the age-spectrum (30–65 years). The percentage of 11A1 labeled astrocytes gradually increases by age (p = 0.020). (E) Number of the astrocytes in measured area across the age-spectrum (30–65 years). Number of astrocytes gradually increases by age (p = 0.014).
