The Immunohistochemical Localization of Glutathione Peroxidase

  • Kiyoshi Akeo
  • Tadahisa Hiramitsu
  • Keiichi Watanabe
Part of the Methods in Molecular Biology™ book series (MIMB, volume 196)


Glutathione peroxidase (GSH-PO), a selenium-dependent and lipid peroxide-scavenging enzyme that effectively reduces lipid peroxides with the concomitant oxidation of glutathione is distributed in mitochondria (1,2). Utsunomiya et al. (3) confirmed the dual localization of GSH-PO in the cytosol and mitochondria of normal rat hepatocytes. We have shown that short-term incubation with linoleic acids (LA) increased the thiobarbituric acid- reactive substance (TBARS) in the RPE cells, which indicated the level of lipid peroxides (4). Mitochondria in the RPE cells were swollen by the incubation with LA or linoleic acid hydroperoxide (LHP) (5). We speculate that exposure of RPE cells to LA or LHP may cause damage to the mitochondria by lipid peroxidation, resulting in the cytotoxicity of RPE cells. We also found loss of mitochondria of bovine RPE cells cultured in hypoxia as low as 1% oxygen, induced malfunction of phagocytosis and a decrease in antioxidants such as glutathione containing sulfur (6).


Sodium Dodecyl Sulfate Laser Scan Microscopy Outer Segment Photoreceptor Cell Subcellular Organelle 
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  1. 1.
    Watanabe, K. (1986) Lipid peroxidation and cell injury. Roles of glutathione peroxidase as a scavenger of lipid peroxidase. Trans. Soc. Pathol. Jpn. 76, 39–74.Google Scholar
  2. 2.
    Zarowski, J. and Tappel, A. L. (1978) Purification and properties of rat liver mitochondrial glutathione peroxidase. Biochem. Biophys. Acta 526, 65–76.Google Scholar
  3. 3.
    Utsunomiya, H., Komatsu, N., Yoshimura, S., Tsutsumi, Y., and Watanabe, K. (1991) Extra ultrastructural localization of glutathione peroxidase in normal rat hepatocytes: advantages of microwave fixation. J. Histochem. Cytochem. 39, 1167–1174.PubMedGoogle Scholar
  4. 4.
    Akeo, K. and Hiramitsu, T. (1998) Changes in lipid peroxide level in retinal pigment epithelial cells in vitro upon addition of linoleic acids or linoleic acid hydroperoxides under varying concentrations of oxygen. Pigment Cell Res. 11, 320–326.PubMedCrossRefGoogle Scholar
  5. 5.
    Akeo, K., Hiramitsu, T., Kanda, T., Yorifuji, H., and Okisaka, S. (1996) Comparative effects of linoleic acid and linoleic acid hydroperoxide on growth and morphology of bovine retinal pigment epithelial cells in vitro. Curr. Eye Res. 5, 467–476.CrossRefGoogle Scholar
  6. 6.
    Akeo, K., Fujiwara, T., Yorifuji, H., and Okisaka, S. (1997) X-ray microanalysis and phagocytotic activity of cultured retinal pigment epithelial cells in hypoxia. Pigment Cell Res. 10, 257–264.PubMedCrossRefGoogle Scholar
  7. 7.
    Poincelot, R. P. and Abrahamson, E. W. (1970) Fatty acid composition of bovine rod outer segments and rhodopsin. Biochim. Biophys. Acta 202, 382–385.PubMedGoogle Scholar
  8. 8.
    Hendricks, T. K., Klompmakers, A. A., Daemen, F. J. M., and Bonting, S. L. (1976) Biochemical aspects of the visual process. XXII. Movement of sodium ions through bilayers composed of retinal and rod outer segment lipids. Biochim. Biophys. Acta 443 271–281.Google Scholar
  9. 9.
    Stone, W. L., Farnsworth, C. C., and Dratz, E. A. (1979) A reinvestigation of fatty acid content of bovine, rat and frog retinal rod outer segments. Exp. Eye Res. 28, 387–397.PubMedCrossRefGoogle Scholar
  10. 10.
    Zigler, J. S. Jr. and Hess, H. H. (1985) Cataracts in the Royal College of Surgeons rat: Evidence for initiation by lipid peroxidation products. Exp. Eye Res. 41, 67–76.PubMedCrossRefGoogle Scholar
  11. 11.
    Doonan, S., Marra E., Passarella S., Saccone C., and Quagliariello, E. (1984) Transport of proteins to mitochondria. Int. Rev. Cytol. 91, 141–186.PubMedCrossRefGoogle Scholar
  12. 12.
    Savanian, A., Mukkassah-Kely, S. F., and Montestruque, S. (1983) The influence of phospholipase A2 and glutathione peroxidase on the elimination of membrane lipid peroxides. Arch. Biochem. Biophy. 223, 441–452.CrossRefGoogle Scholar
  13. 13.
    Itoh, J., Osamura, Y., and Watanabe, K. (1992) Subcellular visualization of light microscopic specimens by laser scanning microscopy and computer analysis: a new application of image analysis. J. Histochem. Cytochem. 40, 955–967.PubMedGoogle Scholar
  14. 14.
    Glick, B. S. (1995) Can Hsp70 proteins act as force-generating motors? Cell 80, 11–14.PubMedCrossRefGoogle Scholar
  15. 15.
    Clarke, I. S., Dzialoszynski, T., Sanford, S. E., and Trevithick, J. R. (1991) A possible relationship between cataract, increased levels of the major heat shock protein Hsp70 and decreased levels of S-antigen in the retina of the RCS rat. Exp. Eye Res. 53, 545–548.PubMedCrossRefGoogle Scholar
  16. 16.
    Yoshimura, S., Komatsu, N., and Watanabe, K. (1980) Purification and immunohistochemical localization of rat liver glutathione peroxidase. Biochem. Biophys. Acta 621, 130–137.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2002

Authors and Affiliations

  • Kiyoshi Akeo
    • 1
  • Tadahisa Hiramitsu
    • 2
  • Keiichi Watanabe
    • 3
  1. 1.Department of OphthalmologyTakasaki National HospitalTakasaki-shiJapan
  2. 2.Photon Medical Research CenterHamamatsu University School of MedicineHamamatsuJapan
  3. 3.Department of PathologyTokai University School of Medicine, Boseidai Isehara-shiKanagawaJapan

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