blood vessels marker cd31  (R&D Systems)

Journal: Alzheimer's & Dementia

Article Title: tPA supplementation preserves neurovascular and cognitive function in Tg2576 mice

doi: 10.1002/alz.13878

Figure Lengend Snippet: Intranasal tPA administration attenuates pial and neocortical CAA in aged Tg2576 mice. A, In vivo two‐photon excited fluorescent images illustrating methoxy‐XO4 + amyloid deposits (magenta) around somatosensory cortex blood vessels (green) identified by retroorbital injection of 70 kDa Texas Red dextran. Methoxy‐XO4 + Aβ deposits are reduced in intranasal tPA‐treated Tg2576 compared to vehicle‐treated Tg2576 mice. B, CAA burden (% thioflavin‐S + area) in cortical sections labeled with thioflavin‐S and the blood vessel marker CD31 is reduced in intranasal tPA‐treated Tg2576 compared to vehicle‐treated Tg2576 mice, but amyloid plaque burden (% thioflavin‐S + plaque area) was comparable between the two groups. In (A) and (B), arrows indicate CAA and arrowheads plaques. N = 5/group; unpaired t test; scale bars, 100 μm in (A) and 200 μm in (B); data presented as mean ± standard error of the mean. Aβ, amyloid beta; CAA, cerebral amyloid angiopathy; tPA, tissue plasminogen activator; WT, wild type.

Article Snippet: To quantify CAA and amyloid plaque burden, , , coronal sections were stained with the blood vessels marker CD31 (goat polyclonal, 1:150, Catalog # AF3628, R&D Systems) followed by thioflavin‐S (0.5%), mounted on slides, and imaged with a confocal microscope (Leica SP8).

Techniques: In Vivo, Injection, Labeling, Marker

Journal: Nmr in Biomedicine

Article Title: Imaging angiogenesis in an intracerebrally induced model of brain macrometastasis using α v β 3 ‐targeted iron oxide microparticles

doi: 10.1002/nbm.4948

Figure Lengend Snippet: (A) Quantitation of CD31 total stained area as a percentage of tumour area in the tumour (blue circles) and the contralateral striatum (red squares), plotted against time with 95% confidence intervals. A significant increase in blood vessels in the tumour is evident after Day 12 (linear regression sum‐of‐squares F‐test, p < 0.05; Spearman's r, tumour regression: 0.75, p < 0.0001; contralateral striatum regression: 0.74, p < 0.0001). (B) Comparison of microvessel density (CD31 positive vessels) in the tumour (blue circles) and the contralateral striatum (red squares), plotted against time with 95% confidence intervals. A significant increase in blood vessels in the tumour is evident after Day 13 (linear regression sum‐of‐squares F‐test, p < 0.05; Spearman's r, tumour regression: 0.56, p < 0.01; contralateral striatum: 0.51, p < 0.05). (C) Quantitation of total α v β 3 ‐stained area as a percentage of tumour area across the timecourse. (D) Quantitation of α v β 3 positive vessels across the timecourse. (E) Number of α v β 3 positive vessels correlated positively with tumour size (Pearson's coefficient p < 0.0001, R 2 = 0.88). (F) Number of α v β 3 positive vessels correlated positively with CD31 microvessel density (Pearson's coefficient, p < 0.001, R 2 = 0.46). In all cases, PBS control ( n = 3), Days 7 ( n = 4), 14 ( n = 6), 21 ( n = 3), 28 ( n = 4), and 35 ( n = 4). Data for these figures had a lognormal distribution, and no outliers were found to be present (ROUT test). Bars represent mean ± standard deviation; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. PBS, phosphate‐buffered saline.

Article Snippet: Brain tissue sections were stained for blood vessel marker CD31 (AF3628, R&D Systems), VCAM‐1 (1510‐14, Southern Biotech), and the macrophage/microglia marker Iba‐1 (ab5076, Abcam, Cambridge, UK), as described previously., , Secondary antibodies used were biotinylated horse anti‐goat IgG (BA‐9500, Vector Laboratories, Peterborough, UK) and biotinylated goat anti‐rat IgG (BA‐9401, Vector Laboratories).

Techniques: Quantitation Assay, Staining, Comparison, Control, Standard Deviation, Saline

Journal: Nmr in Biomedicine

Article Title: Imaging angiogenesis in an intracerebrally induced model of brain macrometastasis using α v β 3 ‐targeted iron oxide microparticles

doi: 10.1002/nbm.4948

Figure Lengend Snippet: Aggregations of iron in 4T1‐GFP tumour tissue. (A) Representative section of gadolinium‐enhancing tumour stained for Perls' Prussian blue (iron, blue) and counterstained with nuclear fast red. Black arrowheads indicate examples of single MPIO associated with the vascular endothelium, and red arrowheads indicate examples of iron aggregations. (B–C) Correlations between iron aggregations and tumour size in mice injected with either (B) RGD‐MPIO or (C) Control RDG‐MPIO. Linear regression analysis showed a positive correlation between number of aggregations and tumour size ( R 2 = 0.64, *** p < 0.001) in mice injected with RGD‐MPIO, but not control RDG‐MPIO, although a similar trend was evident; 95% confidence intervals are shown. (D–F) Consecutive sections stained for (D) Iba‐1 (macrophages and microglia, brown staining), (E) Prussian blue (iron), and (F) CD31 (blood vessels, brown staining) in a Day 35 mouse injected with RGD‐MPIO. The red arrow indicates a larger iron aggregation in a similar location to macrophage staining (D; Iba‐1), while the black arrow indicates single MPIO distant from macrophage staining, but close alignment with a blood vessel (F; CD31). (G–H) Double staining of 4T1‐GFP tumour tissue sections, from a mouse injected with RGD‐MPIO at the Day 35 timepoint, for Prussian blue (iron) and macrophages/microglia (Iba‐1, brown staining), indicate colocalisation of iron within macrophages/microglia (red arrows). Sections counterstained with nuclear fast red. (I) Double staining for Prussian Blue and CD31 revealed single MPIO (black arrow) bound to blood vessels (brown stained) in mice injected with RGD‐MPIO; representative image from Day 21 shown. (J) In the gadolinium‐enhancing 4T1‐GFP tumours, single endothelium‐bound MPIO are observed more often in mice injected with RGD‐MPIO ( n = 10) than control RDG‐MPIO ( n = 3, t ‐test, ** p < 0.01). (K) Histogram showing the cumulative frequency of the measured distance between the centre of the iron‐laden macrophages and the centre of the blood vessel lumen, indicating their close association with blood vessels. Scale bar = 25 μm in (A) and 10 μm in (D–I). MPIO, microparticles of iron oxide; RDG, Arg‐Asp‐Gly peptide, scrambled control; RGD, Arg‐Gly‐Asp peptide, targeting integrin α v β 3 .

Article Snippet: Brain tissue sections were stained for blood vessel marker CD31 (AF3628, R&D Systems), VCAM‐1 (1510‐14, Southern Biotech), and the macrophage/microglia marker Iba‐1 (ab5076, Abcam, Cambridge, UK), as described previously., , Secondary antibodies used were biotinylated horse anti‐goat IgG (BA‐9500, Vector Laboratories, Peterborough, UK) and biotinylated goat anti‐rat IgG (BA‐9401, Vector Laboratories).

Techniques: Staining, Injection, Control, Double Staining