Abstract
Free radicals are central mediators of a variety of cardiovascular diseases. It has been hypothesized that free radical metabolism, oxygenation, and nitric oxide generation in biological organs such as the heart may vary over the spatially defined tissue structure. There has been great interest in the measurement and spatial imaging of in vivo radical generation and metabolism in the heart. EPR spectroscopy can directly measure free radicals; however, it had not been possible to measure important biological radicals in situ in the heart because conventional spectrometers are not suitable for measurements on large aqueous structures such as whole organs or tissues. This chapter summarizes the design, construction, and application of instrumentation for EPR spectroscopy and imaging of the isolated heart. The spectrometer consists of an L-band microwave bridge with the source locked to the resonant frequency of a specially designed loop-gap resonator or reentrant resonator. Radical concentrations as low as 0.2 µM in aqueous solutions could be measured. Studies of isolated beating hearts involving simultaneous real time measurements of free radicals and cardiac contractile function are described. Applications studying the kinetics of free radical metabolism in normally perfused and globally ischemic hearts are reviewed. It is also demonstrated that this technique can be used to noninvasively measure myocardial oxygenation. To address fundamental questions regarding the role of spatially localized alterations in radical metabolism, oxygenation, and nitric oxide in the pathophysiology of cellular injury during ischemia, instrumentation was developed and optimized for 3D spatial and 3D or 4D spectral-spatial imaging of free radicals in the isolated perfused rat heart at 1.2 GHz. High quality 3D spectral-spatial imaging of nitroxide metabolism was performed as well as spatially localized measurements of oxygen concentrations, derived from the oxygen dependent linewidth broadening. In these spectral-spatial images submillimeter resolution was observed enabling visualization of the left ventricular and right ventricular myocardium. With 3D spatial imaging using single-line labels, resolutions down to 100 um were obtained enabling visualization of the ventricles, aortic root, and proximal coronary arteries. Using metal complexes which trap nitroc oxide, measurement and imaging of nitric oxide generation during cardiac ischemia was performed. With the use of 15N isotope labeling it was possible to map the metabolic pathway of this nitric oxide generation. EPR imaging is a powerful tool that can provide unique information regarding the spatial localization of free radicals, oxygen, and nitric oxide in biological organs such as the heart.
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Zweier, J.L., Samouilov, A., Kuppusamy, P. (2003). Cardiac Applications of in Vivo EPR Spectroscopy and Imaging. In: Berliner, L.J. (eds) In Vivo EPR (ESR). Biological Magnetic Resonance, vol 18. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-0061-2_16
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DOI: https://doi.org/10.1007/978-1-4615-0061-2_16
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