Abstract
To date three distinct isoforms of nitric oxide synthase (NOS) have been identified. Two isoforms are considered to be expressed constitutively—neu-ronal NOS (nNOS; type I NOS) and endothelial NOS (eNOS; type III NOS). The third isoform is not generally present in normal cells and tissues but is induced in response to infection, inflammation or trauma—inducible NOS (iNOS; type II NOS). In 1990 Bredt and Synder (1) succeeded in developing antibodies to rat brain NOS (nNOS) and used immuncytochemistry subsequently to furnish one of the first anatomical descriptions of the distribution and localization of nNOS. Today numerous antibodies to all three NOS isoforms isolated from various tissues and different animal species are available and the application of immunocytochemistry is commonplace in the investigation of NOS in healthy and diseased tissues including human (2–6) (Fig. 1 Fig. 2 Fig. 3).
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References
Bredt, D. S., Hwang, P. M., and Snyder, S. H. (1990) Localization of nitric oxide synthase indicating a neural role for nitric oxide. Nature 347, 768–770.
Springall, D. R., Riveros-Moreno, V., Buttery, L. D. K., Suburo, A., Bishop, A. E., Moncada, S., and Polak, J. M. (1990) Immunological detection of nitric oxide synthase(s) in human tissues using heterologous antibodies suggesting different isoforms. Histochemistry 98, 259–266.
Kobzick, L., Bredt, D. S., Lowenstein, C. J., Drazen, J., Gaston, B., Sugarbaker, D., and Stamler, J. S. (1993) Nitric oxide synthase in human and rat lung: immunocytochemical and histochemical localization. J. Histochem. Cytochem. 9, 371–377.
Shaul, P. W., North, A. J., Wu, L. C., Wells, L. B., Brannon, T. S., Lau, K. S., Michel, T., Margraf, L. R., and Star, R. A. (1994) Endothelial nitic oxide synthase is expressed in cultured human bronchiolar epithelium. J. Clin. Invest. 94, 2231–2236.
Tracey, W. R., Xue, C., Klinhhofer, V., Barlow, J., Pollock, J. S., Forstermann, U., and Johns, R. A. (1994) Immunological detection of inducible NO synthase in human lung. Am. J. Physiol. 266, L722–727.
Nicholson, S., Bonecini-Almeida, Lapa e Silva, J. R., Nathan, C., Kie, Q. W., Mumford, R., Weidner, R., Calagcay, J., Geng, J., Boechat, N., Linhares, C., Rom, W., and Ho, J. L. (1996) Inducible nitric oxide synthase in pulmonary alveolar macrophages from patients with tuberculosis. J. Exp. Med. 183, 2293–2302.
Polak, J. M. and Van Noorden, S., eds. (1997) Introduction to Immun-cytochemistry, BIOS Scientific Publishers, Oxford, UK.
Pollock, J. S., Nakane, M., Buttery, L. D. K., Martinez, A., Springall, D. R., Polak, J. M., Forstermann, U., and Murad, F. (1993) Characterization and localization of endothelial nitric oxide synthase using specific monoclonal antibodies. Am. J. Physiol. 265, C1379–C1387.
Buttery, L. D. K., McCarthy, A., Springall, D. R., Sullivan, M. H. F., Elder, M. G., Michel, T., and Polak, J. M. (1994) Endothelial nitric oxide synthase in the human placenta: regional distribution and proposed regulatory role at the feto-maternal interface. Placenta, 15, 257–265.
Buttery, L. D. K., Chester, A. H., Springall, D. R., Borland, J. A. A., Michel, T., Yacoub, M. H., and Polak, J. M. (1996) Explanted vein grafts with an intact endothelium demonstrate reduced focal expression of endothelial NO synthase specific to atherosclerotic sites. J. Pathol. 179, 197–203.
Buttery, L. D. K., Springall, D. R., Chester, A. H., Evans, T. J., Standfield, N., Parums, D. V., Yacoub, M. H., and Polak, J. M. (1996) Inducible NO synthase is present within human atherosclerotic lesions and promotes the formation and activity of peroxynitrite. Lab. Invest. 75, 77–85.
Higman, D. J., Strachan, A. M. J., Buttery, L. D. K., Hicks, R. C. J., Springall, D. R., Greenhalgh, R. M., and Powell, J. T. (1996) Smoking impairs the activity of endothelial nitric oxide synthase in saphenous vein. Arterioscler. Thromb. Vasc. Biol. 16, 546–552.
Evans, T. J., Buttery, L. D. K., Carpenter, A., Springall, D. R., Polak, J. M., and Cohen J. (1996) Inducible nitric oxide synthase is localized within primary granules of cytokine-treated human neutrophils and produces peroxynitrite-mediated nitration of ingested bacteria. Proc. Natl. Acad. Sci. 93, 9553–9558.
Coers, W., Timens, W., Kempinga, C., Klok, P., and Moshage, H. (1998) Specificity of antibodies to nitric oxide synthase isoforms in human, guinea pig, rat and mouse tissues. J. Histochem. Cytochem. 46, 1385–1391.
Mason, N. A., Springall, D. R., Burke, M., Pollock, J., Mikhail, G., Yacoub, M. H., and Polak, J. M. (1998) High expression of endothelial nitric oxide syn-thase in plexiform lesions of pulmonary hypertension. J. Pathol. 185 313–318.
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Buttery, L.D.K., Polak, J.M. (2000). Immunochemical Detection of Nitric Oxide Synthase in Human Tissue. In: Evans, T.J. (eds) Septic Shock Methods and Protocols. Methods in Molecular Medicine™, vol 36. Humana Press. https://doi.org/10.1385/1-59259-216-3:145
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DOI: https://doi.org/10.1385/1-59259-216-3:145
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