In vivo ESR measurement of free radical reaction in living mice

Recently, free radicals such as active oxygen species, nitric oxide, etc are believed to be one of the key substances in physiological and pathological, toxicological phenomena, and oxidative damages, and all organism have defencing system against such as free radicals. Formation and extinction of free radicals may be regulated through bio-redox system, in which various enzymes and compounds should be involved in very complicated manner. Thus, direct and non-invasive measurement of in vivo free radical reactions with living animals must be essential to understand the role of free radicals in pathophysiological phenomena. Electron spin resonance spectroscopy (ESR) is very selective and sensitive technique to detect free radicals, but a conventional ESR spectrometer has large detect in application to living animals, since high frequent microwave is absorbed with water, resulting in generation of high fever in living body. In order to estimate in vivo free radical reactions in living whole animals, we develop in vivo ESR-CT technique using nitroxide radicals as spin probes. Nitroxide radicals and their reduced forms, hydroxylamines, are known to interact with various redox systems. We found that! ! the signal decay due to reduction of nitroxyl radicals is influenced by aging, inspired oxygen concentration, ischemia-referfusion injury, radiation, etc. In the present paper, I will introduce in vivo ESR technique and my laboratory recent results concerning non-invasive evaluation of free radical reactions in living mice.