Abstract
It is well known that the superoxide anion radical (O-2) is produced in the body as a result of biochemical activities of many cells, such as neutrophils, macrophages, and Kupffer cells (1). Production of O-2 is detectable by various methods, including cytochrome-c oxidation (1), nitroblue tetrazolium (NBT) staining, and chemiluminescence (CL) measurement (2). The site and amount of O-2 production is variable with time and readily determined in vivo by applying the CL method. In earlier studies, the CL method was successfully applied for detection of O-2 generation at a tissue level in the lung (3), liver (4), pancreas (5), heart (6), and brain (7). Combined with video imaging techniques, the CL method makes it possible to analyze O-2 generation at a high spatial resolution, sometimes even at a single cell level. In this chapter, we describe the CL method applied to O-2 detection from the islets of Langerhans in the perfused pancreas.
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Halliwell, B. and Gutteridge, J. M. C. (eds.) (1989) Free Radicals in Biology and Medicine, 2nd ed. Clarendon Press, Oxford.
Nakano, M. (1998) Detection of active oxygen species in biological systems. Cell Mol. Neurobiol. 18, 565–579.
Takahashi, A., Nakano, M., Mashiko, S., and Inaba, H. (1990) The first observation of O-2 generation in situ lungs of rats treated with drugs to induce experimental acute respiratory distress syndrome. FEBS Lett. 261, 369–372.
Nakano, M., Kikuyama, M., Hasegawa, T., Ito, T., Sakurai, K., and Hashimura, E. (1995) The first observation of O-2 generation at real time in vivo from nonKupffer sinusoidal cells in perfused rat liver during acute ethanol intoxication. FEBS Lett. 372, 140–143.
Kishimoto, W., Nakao, A., Nakano, M., Takahashi, A., Inaba, H. and Takagi, H. (1995) Detection of superoxide free radicals in rats with acute pancreatitis. Pancreas 11, 122–126.
Ushiroda, S., Maruyama, Y., and Nakano, M. (1997) Continuous detection of superoxide in situ during ischemia and reperfusion in the rabbit heart. Jpn. Heart J. 38, 91–105.
Yamamoto, S., Terakawa, S., Sakurai, T., Matsumura, S., and Uemura, K. (1997) Production of reactive oxygen species and glutamate neurotoxicity in a rat organotypic hippocampal culture. Adv. Neurotrauma Res. 9, 61–64.
Sakurai, T. and Terakawa, S. (1997) Radical production and degranulation in pancreatic islets induced by glucose. Jpn. J. Physiol. 47 (Suppl. 1), S15.
Mizuguchi, Y. (1998) Single photon counting, in Methods in Molecular Biology, vol. 108: Free Radical and Antioxidant Protocols (ed.), Humana Press, NJ, pp. 37–55.
Suzaki, E., Kobayashi, H., Kodama, Y., Masujima, T., and Terakawa, S. (1997) Video-rate dynamics of exocytotic events associated with phagocytosis in neutrophils. CellMotil. Cytoskeleton 38, 215–228.
DiGregorio, K. A., Cilento, E. V., and Lantz, R. C. (1987) A kinetic model of superoxide production from single pulmonary alveolar macrophages. Am. J. Physiol. 252, C677–C683.
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© 2002 Humana Press Inc.
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Sakurai, T., Terakawa, S. (2002). Superoxide Production in the Islet of Langerhans Detected by the MCLA Chemiluminescence Method. In: Armstrong, D. (eds) Oxidants and Antioxidants. Methods in Molecular Biology™, vol 196. Humana Press. https://doi.org/10.1385/1-59259-274-0:203
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DOI: https://doi.org/10.1385/1-59259-274-0:203
Publisher Name: Humana Press
Print ISBN: 978-0-89603-851-6
Online ISBN: 978-1-59259-274-6
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