Fig. 14
From: Spatiotemporal diversity in molecular and functional abnormalities in the mdx dystrophic brain
Histochemical analysis of the distribution of blood vessels and microglia in C57Bl/10 (w/t) and mdx brain ATPase/CD39 activity was performed in situ by incubating cryosections of A w/t and B mdx mouse brains with 300 μM ATP in the presence of Pb(NO3)2, followed by microscopic detection of ATP-derived inorganic phosphate as a brown precipitate. TNAP activity was measured in C w/t and D mdx mouse brains using the chromogenic substrates BCIP and NBT and subsequent monitoring of the development of blue colour reactions. E LMA-embedded brain sections from w/t (upper panels) and mdx (lower panels) mice were stained for the molecular microglia markers Iba1, P2Y12R and CD39 (with the latter enzyme also co-expressed on vascular endothelial cells), as indicated. 3D images of 100 µM tissue were captured using a spinning disk confocal microscope. Maximum intensity projections for each channel are shown in grayscale, with the right panels displaying merged images with nuclei counterstained with DAPI. BV blood vessel, CC corpus callosum, Cereb cerebellum, FL frontal lobe, MG microglia, Thal thalamus. Scale bars: 2 mm (A), 300 μm (insets in A, B), and 80 μm (E)