Figure 2.
Temporal and regional distribution of the regulators of microglial proliferation during prion disease. A, Analysis of the expression of mRNA of CSF1, IL34, and CSF1R in the hippocampus (CA1) and thalamus of prion disease (ME7) and control (NBH) mice. Expression of CSF1, IL34, and CSF1R is represented as mean ± SEM and indicated as relative expression levels using the 2ΔΔCt method. B, C, F–H, Immunohistochemical analysis of the expression of IL34 (B, C), PU.1 (F, G), and C/EBPa (F, H) in the CA1 region of the hippocampus (see representative images) and the thalamus of prion disease (ME7) and control (NBH) mice. Quantified data expressed as mean ± SEM of the number of IL34+, PU.1+, or C/EBP+ cells per square millimeter. D, Analysis of the expression of IL34 in glial cells by triple immunofluorescence for IL34 (red), GFP (microglia, green), and GFAP (astrocytes, blue) in the hippocampus (CA1) of prion disease mice (ME7). E, Analysis of the expression of mRNA of PU.1, GATA1, and C/EBPa in the hippocampus (CA1) and thalamus of prion disease (ME7) and control (NBH) mice. Expression of PU.1, GATA1, and C/EBPa is represented as mean ± SEM and indicated as relative expression levels using the 2ΔΔCt method. I, Analysis of the expression of PU.1 or C/EBPa (red) in microglial cells (GFP, green) by double immunofluorescence in the hippocampus (CA1) of prion disease mice (ME7). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Data were analyzed with a two-way ANOVA and a post hoc Tukey test (n = 6). C, G, H, Nuclei are stained with H/E (blue). I, Nuclei are stained with Hoechst (blue). D, I, Fluorescent sections evaluated with confocal microscopy. Scale bars: C, G, H, 100 μm; insets, 50 μm; D, I, 50 μm.