Fig. 2

DNA damage does not accompany arrested C-like-mitoses. a-d DNA fragmentation detected with TUNEL is not typical for astrocytes arrested in mitosis. a CA1 hippocampal subfield with many mitotic astrocytes (identified with phospho-vimentin (pVIM), some of the astrocytes arrested in metaphase are indicated with arrows) that are not positive for TUNEL. Note that the TUNEL+ green nuclei are located in the pyramidal layer and belong to neurons. b, c Piriform cortex, astrocyte with many small nuclei (arrow in b) and astrocyte arrested in metaphase (arrow in c) do not show TUNEL staining. Note that many nuclei are TUNEL+, but are not astrocytes. d TUNEL+ staining of misaligned metaphase chromosomes indicating DNA fragmentation in this astrocyte (arrow). Hippocampus, CA1 subfield. Insets in lower right parts in (b, c, and d) show enlarged nuclei and metaphase chromosomes in the cells marked with arrows in (b, c, d) respectively. e-g Immunoreactivity for phospho-gamma-H2AX (H2AX) is present only in small numbers of mitotic astrocytes. e H2AX+ nuclei (arrows) in the CA1 layer of pyramidal neurons. Note in E that many neurons displayed shrunken, condensed nuclei indicating apoptotic changes. e1, e1’) enlarged boxed area in E shows mitoses. Note that positive signals for H2AX are localized in small, focal areas of chromosomes. f, g Immunoreactivity for H2AX is present in some metaphase chromosomes (f, arrow) and in micronuclei (g, arrow) indicating DNA breaks. h, i, j Colocalization of H2AX with markers of kinetochores: BubR1 (i), Aurora B (j), and survivin (k). f-j – piriform cortex. All images were obtained with confocal microscope 4 days after pilocarpine administration. Scale bars:  a = 150 μm,  b-d = 35 μm,  e = 125 μm,  f,g = 8 μm, h-j = 3 μm