Figure 8.
Lateral fluid percussion TBI induces protein nitration in the rat brain. A, H+E-stained coronal section demonstrating gross histological damage (arrow) 7 d after fluid percussion injury. B, Low-power view (phase contrast) of the area of cortical injury (24 h) after lateral fluid percussion injury in the rat. The dotted line outlines the injury area. Scale bar, 500 μm. C, Anti-NT staining of same section, indicating increased NT staining (arrow) in the injured area. Immunoreactivity was visualized using DAB, and sections were counterstained using cresyl violet. A–C are representative of at least six animals per experiment. D, Representative brain sections from sham and injured animals stained with TTC to delineate the injury site (arrow) for protein harvest. E, Immunoblots using anti-3-nitrotyrosine antibodies from injured (I) and contralateral (C) cortex taken 24 h after TBI or control (sham) animals (n = 6 animals/experiment). Notably, this monoclonal antibody (4 μg/ml; BioMol) detects this ∼30 kDa band more selectively than the polyclonal antibody (Upstate Biotechnology) used in the tissue culture experiments (Fig. 2D). F, Immunoblot of caspase-3 (both pro- and active/cleaved forms) from brain lysates taken at 24 h from cortex ipsilateral (I) and contralateral (C) to FPI in four animals subjected to a 2.4 atm injury. Positive control, 20 μg recombinant human caspase-3 (Sigma).