Figure 2

Pathology in the striatum and hippocampus of adult ischemia and hypoxia (HI) mice. (A) Overview of the pathology in striatum and hippocampus of sham and HI mice at 4 weeks after HI conditions. Obvious neuron loss (stained with a neuronal marker, NeuN, green) and reactive astrocytes (stained with an astrocytic marker, GFAP, red) are in these two areas. (B,C) The magnified images of the injured area in striatum (B) and hippocampus (C). (D) Quantification data of the injured area, calculated as the GFAP⁺ overexpressed area ratio with the total area of the striatum and hippocampus. N = 5 mice with 4 slices per animal. Abbreviations: Str, striatum; HP, hippocampus. (E) showing the scarce expression of another neuronal marker, MAP2 (green), on the neuronal dendrites in the ischemic CA1 and CA3 areas, indicating the disruption of synaptic connections. (F) Represent the neuron degeneration by FJB⁺ cells (green) in ischemic striatum and CA1 area 24 h after HI conditions. (G)–(J) Data showed the morphological changes of astrocytes in HI mice 4 weeks after stroke. (G,I) Quantification of the significant increase in GFAP⁺ expression in HI group than sham. Unpaired t-test, **P < 0.01, ***P < 0.001. N = 3 mice in each group with 3 slices per animal. Data are collected from three fields in striatum and one field in each hippocampal sub-region shown in cartoons. (H,J) Fluorescent sample images and cropped cells with binary images showed the hypertrophy status of reactive astrocytes by larger cell area in the ischemic striatum (H) and CA1 (J). Unpaired t-test, **P < 0.01, ***P < 0.001. For area analysis, N = 15 cells in each slice with 3 slices per animal. The images were all captured by the fluorescent microscope (Nikon Eclipse Ni). The cropped cells with binary images in (H,J) were performed in Image J 2.0.0 (FIJI). Quantitative analysis of the fluorescence intensity and morphological changes were performed in FIJI. Data are shown as the mean and SEM. Scale bars in (A) represent 500 μm, (F) 100 μm and (J) 10 μm.