Summary
Glutathione in tissues forms an intense fluorophore with a solution ofo-phthalaldehyde at room temperature. We have studied the loss of glutathione from tissue sections and find that it is not measurable from thick sections. The fluorescence spectra of the induced fluorophore between glutathione ando-phthalaldehyde are identical in model and tissue sections, while depletion of hepatic glutathione by diethyl maleate produces a comparable fall in fluorescence measured biochemically or histochemically. This simple method is specific as interfering substances, such as spermine and spermidine, produce very weak fluorescence under the conditions employed.
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ADAMS, D. J., CARMICHAEL, J. & WOLF, C. R. (1985) Altered mouse bone marrow glutathione and glutathione transferase levels.Cancer Res. 45, 1669–73.
ASGHAR, K., REDDY, B. G. & KRISHNA, G. (1975) Histochemical localization of glutathione in tissues.J. Histochem. Cytochem. 23, 774–9.
CASINI, A. F., POMPELLA, A. & COMPORTI, M. (1985) Liver glutathione depletion induced by bromobenzene, iodobenzene, and diethyl maleate poisoning and its relation to lipid peroxidation and necrosis.Am. J. Pathol. 118, 225–37.
CHIECO, P. & BOOR, P. J. (1983) Use of low temperatures for glutathione histochemical stain.J. Histochem. Cytochem. 31, 975–6.
COHN, V. H. & LYLE, J. (1966) A fluorometric assay for glutathione.Anal. Biochem. 14, 434–40.
CROSS, S. A. M., EWEN, S. W. B. & ROST, F. W. D. (1971) A study of the methods available for the cytochemical localization of histamine by fluorescence induced witho-phthalaldehyde or acetaldehyde.Histochem. J. 3, 471–6.
HISSIN, P. J. & HILF, R. (1976) A fluorometric method for determination of oxidized and reduced glutathione in tissues.Anal. Biochem. 74, 214–26.
HOUGAARD, D. M. & LARSSON, L. I. (1982) Polyamine cytochemistry. Use of a novelo-phthalaldehyde method for visualizing spermidine and spermine. Comparisons to the formaldehyde—fluorescamine method.Histochemistry 76, 247–59.
JOLLOW, D. J., MITCHELL, J. R., ZAMPAGLIONE, N. & GILLETTE, J. R. (1974) Bromobenzene-induced liver necrosis. Protective role of glutathione and evidence for 3,4 bromobenzene oxide as the hepatotoxic metabolite.Pharmacology 11, 151–69.
KAPLOWITZ, N., AW, T. K. & OOKHTENS, M. (1985) The regulation of hepatic glutathione.Ann. Rev. Pharmacol. Toxicol. 25, 715–44.
MEISTER, A. & ANDERSON, M. E. (1983) Glutathione.Ann. Rev. Biochem. 52, 711–60.
MOKRASCH, L. C. & TESCHKE, E. J. (1984) Glutathione content of cultured cells and rodent brain regions: a specific fluorometric assay.Anal. Biochem. 140, 506–9.
ORRENIUS, S. & MOLDEUS, P. (1984) The multiple roles of glutathione in drug metabolism.Trends Pharm. Sci. 5, 432–5.
PEARSE, A. G. E. (1985)Histochemistry: Theoretical and Applied, Vol. 2, 4th edn., p. 483. Edinburgh, London, New York: Churchill Livingstone.
SMITH, M. T., LOVERIDGE, N., WILLS, E. D. & CHAYEN, J. (1979) The distribution of glutathione in the rat liver lobule.Biochem. J. 182, 103–8.
TREUMER, J. & VALET, G. (1986) Flow cytometric determination of glutathione alterations in vital cells byo-phthalaldehyde (OPT) staining.Exp. Cell. Res. 163, 518–24.
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Murray, G.I., Burke, M.D. & Ewen, S.W.B. Glutathione localization by a novelo-phthalaldehyde histofluorescence method. Histochem J 18, 434–440 (1986). https://doi.org/10.1007/BF01675336
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DOI: https://doi.org/10.1007/BF01675336