Journal:

Article Title: Recombinant rabies virus vaccine strain SAD-L16 inoculated intracerebrally in young mice produces a severe encephalitis with extensive neuronal apoptosis

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Figure Lengend Snippet: Transferase-mediated dUTP-biotin nick end labelling (TUNEL) and staining (A to D) and immunoperoxidase staining for activated caspase-3 (E to H) in L16-infected mouse brains. The TUNEL staining is present in neurons in the cerebellar external and internal granular layers (A) 6 d postinoculation (pi), in many neurons in the cerebral cortex (B) 4 d pi, and in hippocampal pyramidal neurons and neurons in the dentate gyrus of the hippocampus (C) 5 d pi, while there is low background staining in the hippocampus of a mock-infected mouse (D). Activated caspase-3 staining is present in many neurons in the internal granular layer of the cerebellum (E) 5 d pi, in some Purkinje cells (F) 4 d pi, in both hippocampal pyramidal neurons and in neurons in the dentate gyrus of the hippocampus (G) 4 d pi, and there is low background staining in the hippocampus of a mock-infected mouse (H). A to D, TUNEL staining — methyl green; E to H, Immunoperoxidase — hematoxylin. Magnification: A × 150; B × 95; C, D × 25; E × 130; F × 390; G, H × 20.

Article Snippet: Sections were successively treated with 5% normal goat serum, rabbit polyclonal antibody directed against cleaved (activated) caspase-3 (Asp175) (Cell Signaling Technology, Beverley, Massachusetts, USA) diluted 1:400 in 2% normal goat serum, biotinylated goat anti-rabbit IgG (Vector Laboratories) diluted 1:100 in 2% normal goat serum, 1% hydrogen peroxide in methanol, avidin-biotinylated horseradish peroxidase complex (Vector Laboratories), 3,3-diaminobenzidine tetrachloride (Polysciences) with 0.01% hydrogen peroxide, and 0.5% cupric sulfate in 0.15 M sodium chloride.

Techniques: TUNEL Assay, Staining, Immunoperoxidase Staining, Infection