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Effect of TMPZ on nitrotyrosine expression in ischemic cerebral regions The formation of nitrotyrosine in the brain was detected as described previously [1] with some modifications. In brief, following brain reperfusion, each post-anesthetic rat was perfusion-fixed with a 4 % paraformaldehyde solution. Brains were removed and dehydrated with 30 % sucrose. Samples were coronally cut into serial 10-µm-thick sections (0.3 mm from the bregma) for immunohistofluorescent staining. Non-specific binding sites were blocked with 2 % BSA, then sections were incubated for 2 h with a primary anti-nitrotyrosine mAb (1:50; Cayman). Sections were then incubated for 1 h with a FITC-conjugated secondary antibody (1:100; Amersham). Areas of the cortex of brain sections were captured using a digital color CCD camera (CoolSNAP-Pro cf) and Image-Pro Plus software 4.5. (Media Cybernetics, Silver Spring, MD). The nitrotyrosine level was evaluated as a marker of peroxynitrite accumulation in the penumbra of brain ischemic regions. Peroxynitrite is generated by a combination of NO and superoxide in the ischemic penumbra [2]. This stud