Journal: Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism

Article Title: The pathologic cascade of cerebrovascular lesions in SHRSP: is erythrocyte accumulation an early phase?

doi: 10.1038/jcbfm.2011.122

Figure Lengend Snippet: Figure 3 STL-, laminin-, and IgG-immunoreactive vessels and ratio of the densities of IgG- and STL-positive vessels in SHRSP with different ages. STL (green)- and IgG (red)-positive vessels are shown in the hippocampus of 24-week-old (A) and 36-week-old (B, D, and E) SHRSP. The small number of IgG-immunoreactive vessels in the younger animal (panel A, 24 weeks, white arrows) and its increase with age (panel B, 36 weeks) must be noted. Panel D shows the histologic magnification in different z-planes of the white rectangle in panel B. In panel C, the ratio of the densities of IgG- and STL-positive vessels in SHRSP is plotted over the different ages of the 9 investigated animals. The result of every counting box is given by a marker in the plot. The resulting parabolic regression function for all regions is depicted by the line. The significant increase of the ratio over the time in all of the investigated regions (P = 0.008) showing an increasing blood–brain barrier disturbance in older SHRSP. In panels E to G, confocal analysis illustrates that IgG (red) is deposited most likely between the endothelium (STL in green) and the basal membrane (Lam-laminin in pink). However, in certain vessel IgG, i.e., plasma is also entering the parenchyma (panel G, white arrow). A thrombus is shown in panel H; both IgG and laminin patterns illustrate the rupture of the vessel wall, suggesting that a bleeding had preceded vessel occlusion. SHRSP, spontaneously hypertensive stroke-prone rat; STL, solanum tuberosum lectin.

Article Snippet: Solanum tuberosum lectin (Vector Laboratories, Burlingame, CA, USA) is a fluorescein-labeled potato lectin containing B50% carbohydrate, mainly composed of arabinose and galactose (Vector Laboratories (Van Damme et al, 2004)).

Techniques: Marker, Membrane, Clinical Proteomics

Journal: Acta Neuropathologica Communications

Article Title: Spontaneous white matter damage, cognitive decline and neuroinflammation in middle-aged hypertensive rats: an animal model of early-stage cerebral small vessel disease

doi: 10.1186/s40478-014-0169-8

Figure Lengend Snippet: Brain leukocyte subpopulations differ between SHR and WKY. Brains of SHR and WKY contained comparable numbers of CD45 high leukocytes while the composition of subpopulations was strongly different (A) . NK cells constituted the predominant immune cell type in SHR. By contrast, the proportion of T cells and monocytes/macrophages (Mo/Mɸ) was higher in WKY. Absolute counts of Mo/Mɸ were decreased (B) whereas NK cells were significantly increased in brains of SHR (C) . Immunofluorescence analysis revealed that CD161 positive NK cells (green) were exclusively located in the brain microvasculature (visualized by solanum tuberosum lectin, STL; red) of SHR. Data are mean ± SD. *p < 0.05 by t-test for n = 3 animals/group, scale bar: 10 μm. PMN, polymorphonuclear granulocytes.

Article Snippet: For immunohistochemistry, 25 μm sections were blocked with 5% goat serum, 0.3% Triton X-100 and PBS for 60 min, and incubated with either rabbit anti-Iba1 (Ionized calcium binding adaptor molecule 1; 1:200; Wako Chemicals, Neuss, Germany), biotinylated solanum tuberosum lectin (STL; 1:300; Linaris, Dossenheim, Germany), mouse anti-15-16A1 (1:500; Hycult Biotech, Beutelsbach, Germany; for T cells), mouse anti-GFAP (1:500, Sigma), mouse anti-CD161 (1:100, NovusBio, Cambridge, UK) for 24 h at 4°C.

Techniques: Immunofluorescence

Journal: Acta Neuropathologica Communications

Article Title: Spontaneous white matter damage, cognitive decline and neuroinflammation in middle-aged hypertensive rats: an animal model of early-stage cerebral small vessel disease

doi: 10.1186/s40478-014-0169-8

Figure Lengend Snippet: T cell compartmentalization and morphology is different in SHR. Absolute T cell counts were significantly lower in brains of SHR (A) . Histological analysis of T cell distribution in both strains revealed that only 20% of total T cells in SHR resided in the meninges and the choroid plexus (CP), while the remaining cells were located in parenchymal microvessels (B) . This distribution was reversed in WKY. (B) shows representative sections of the meninges in SHR and WKY. T cells were identified by the pan T cell marker 15-16A1 (green). Distinct T cell compartmentalization was accompanied by a decrease of IL-10, but similar levels of IFNγ in the cerebrospinal fluid (CSF) of SHR (C) . Numbers of intravascular T cells neither differed in the total brain nor in the deep cortical region (DCR) (D) . In SHR, two-thirds of all T cells were rod-shaped while the rod-to-round ratio was almost 1:1 in WKY (D) . Representative images of rod and round shaped T cells (green) in brain microvessels (visualized by solanum tuberosum lectin, STL; red). mRNA expression of vascular adhesion molecule 1 (VCAM-1) was significantly increased in brain endothelial cells of SHR (E) . Data are mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 by t-test for n = 3 (A) , n = 4 (B, C and D) or n = 5 (E) animals/group, scale bar: (B) 20 μm; (D) 5 μm.

Article Snippet: For immunohistochemistry, 25 μm sections were blocked with 5% goat serum, 0.3% Triton X-100 and PBS for 60 min, and incubated with either rabbit anti-Iba1 (Ionized calcium binding adaptor molecule 1; 1:200; Wako Chemicals, Neuss, Germany), biotinylated solanum tuberosum lectin (STL; 1:300; Linaris, Dossenheim, Germany), mouse anti-15-16A1 (1:500; Hycult Biotech, Beutelsbach, Germany; for T cells), mouse anti-GFAP (1:500, Sigma), mouse anti-CD161 (1:100, NovusBio, Cambridge, UK) for 24 h at 4°C.

Techniques: Marker, Expressing