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Electron paramagnetic resonance imaging (EPRI) is a new modality for visualizing O(2) distribution in tissues, such as the brain after stroke or after administration of drugs of abuse. We have recently shown that 3-acetoxymethoxycarbonyl-2,2,5,5-tetramethyl-1-pyrrolidinyloxyl [1] is a pro-imaging agent that can cross the blood-brain barrier. After hydrolysis by esterases, the anion of 3-carboxy-2,2,5,5-tetramethyl-1-tetramethyl-1-pyrrolidinyloxyl [2] is trapped in brain tissue. In this study, we investigated the feasibility of using this to map the changes of O(2) concentration in mouse brain after focal ischemia. The decrease in tissue O(2) concentration in the ischemic region of mouse brain was clearly visualized by EPRI. The hypoxic zone mapped by EPRI was spatially well correlated with the infarction area in the brain imaged by diffusion-weighted magnetic resonance imaging (MRI). Finally, we observed a decrease in the size of the hypoxic region when the mouse breathed higher levels of O(2). This finding suggests that EPRI with specifically designed nitroxides is a promising imaging modality for visualizing O(2) distribution in brain tissue, especially in an ischemic brain. We believe that this imaging method can be used for monitoring the effects of therapeutic intervention aimed at enhancing brain O(2) supply, which is crucial in minimizing brain injury after stroke.

Direct Visualization of Mouse Brain Oxygen Distribution by Electron Paramagnetic Resonance Imaging: Application to Focal Cerebral Ischemia