Fig. 2. Increased extravasation of leukocytes in the brain parenchyma of naïve pericyte-deficient brain. (A) Overview images of the periventricular areas in the brains of control and pericyte-deficient mice (Pdgfbret/ret). Arrowheads indicate leukocyte infiltrates (CD45, in red) in the parenchyma of Pdgfbret/ret mice. Blood vessels are visualized by collagen IV staining (in green). (B) High-magnification images showing a parenchymal infiltrate of CD45hi leukocytes (in red, asterisk) in the corpus callosum of Pdgfbret/ret mice. In control mice, few CD45hi leukocytes (arrowheads) are found only in the lumen of blood vessels (podocalyxin, in green). (C) Quantification of CD45hi leukocytes in different anatomical regions in the brains of control and Pdgfbret/ret mice (n = 3). (D) Quantification of extravasated CD45hi leukocytes in different anatomical regions in the brains of control and Pdgfbret/ret mice (n = 3). One-way ANOVA followed by Tukey’s post hoc test was used to determine the statistical significance. (E) Representative flow-cytometry pseudocolor plots showing the manual gating of microglia and other immune cell populations in the brain of naïve control and Pdgfbret/ret mice. (F) Quantification of the absolute cell numbers of detected immune cell populations using flow cytometry in the brain (n = 7–8 mice per genotype). Pooled data from two independent experiments. Statistical significance was determined using unpaired t test (DCs, CD11c+, MHC-IIhigh, and CD4+ and CD8+ T cells) or Mann–Whitney U test (CD45hi cells, CD45hiCD11b+ cells, MdCs, Ly6Chi monocytes, neutrophils, and B cells). Data are presented as the mean ± SD. (Scale bars: A, 250 μm; B, 100 μm; Insets, 20 μm.)
...read more