Journal: bioRxiv

Article Title: Arteriolar degeneration and stiffness in cerebral amyloid angiopathy are linked to β-amyloid deposition and lysyl oxidase

doi: 10.1101/2024.03.08.583563

Figure Lengend Snippet: Atomic force microscopy (AFM) and the stiffness of arterioles. (a) Coronally sectioned profiles of arterioles from cases with CAA and without CAA were stained with thiazine red and DAPI. Cross-sections of the vessel wall were evaluated by AFM to determine tissue stiffness (example density maps across the vessel wall are shown for each condition). (b) Average stiffness in kPa for non-CAA (40.81±7.19) and CAA cases (157.4 ±6.25) are shown. (c) Increased tissue stiffness could be partly mediated by the activity of extracellular matrix crosslinking enzymes like lysyl oxidase (LOX). Confocal microscopy images of arterioles stained with collagen IV-488 (green), LOX (magenta), and β-amyloid staining with methoxy X04 (blue) shows substantial LOX deposition in CAA, especially in degenerating arterioles (rightmost image).

Article Snippet: Floating sections of post-mortem human tissue were prepared on a Leica 1200 S vibratome with 50 μm and 100 μm thickness. β-amyloid was detected using thiazine red (1 µM, Cat# 2150-33-6, Chemsavers) associated with DAPI (0.5 µM, Cat# 1485, Vector lab) and a mouse monoclonal antibody against VSM α-smooth muscle actin (Cat# ABT1487, Millipore), associated with corresponding secondary antibody, Alexa Fluor 488 (Thermo Fisher A21206).

Techniques: Microscopy, Staining, Activity Assay, Confocal Microscopy

Journal: bioRxiv

Article Title: Arteriolar degeneration and stiffness in cerebral amyloid angiopathy are linked to β-amyloid deposition and lysyl oxidase

doi: 10.1101/2024.03.08.583563

Figure Lengend Snippet: Association of β-amyloid accumulation in the wall of arteriole and structural degeneration and rupture. ( a) Two-dimensional staining of β-amyloid with Thiazine red (red) and nuclei in DAPI (blue). (b) Spearman correlation between vascular β-amyloid surface reconstruction and VSM. (c) Two-dimensional staining of β-amyloid accumulation on arterioles and the absent staining of VSM in local β-amyloid accumulation. The arrow in the upper middle image shows VSM outside the tunica media consistent with a fibrotic process. The arrow in the lower middle image indicates the site of a focal stricture in the vessel wall.(d) Spearman correlation between vascular β-amyloid and average diameter, variability of diameter, and tortuosity. E. Three-dimensional images of ruptured arterioles with staining of β-amyloid with Thiazine red (red) and Collagen IV-488; the first two images show native fluorescence, the last two show vascular surface rendering. Disorganized ”shards” of β-amyloid are present near the site of each rupture. Detailed views of these ruptured vessels can be seen in supplementary video 4.

Article Snippet: Floating sections of post-mortem human tissue were prepared on a Leica 1200 S vibratome with 50 μm and 100 μm thickness. β-amyloid was detected using thiazine red (1 µM, Cat# 2150-33-6, Chemsavers) associated with DAPI (0.5 µM, Cat# 1485, Vector lab) and a mouse monoclonal antibody against VSM α-smooth muscle actin (Cat# ABT1487, Millipore), associated with corresponding secondary antibody, Alexa Fluor 488 (Thermo Fisher A21206).

Techniques: Staining, Fluorescence