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Immunoelectron microscopy of Rosenthal fibers

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Summary

Seventeen intracerebral gliomas containing Rosenthal fibers (RF) were studied by an immunoperoxidase method for localization of ubiquitin (UB), glial fibrillary acidic protein (GFAP), desmin and vimentin (VIM). The majority of RF showed an immunohistochemically negative core surrounded by a ring of overlapping reactions for UB, GFAP and VIM. Many RF were entirely negative for UB and intermediate filaments (IF). Immunoelectron microscopic lozalization of UB and GFAP was performed on seven selected tumors. UB was found in all RF and on IF in the proximity of RF. GFAP reaction was localized on astrocytic IF, including those trapped within RF, and within the granular component of some RF. In contrast to the light microscopic studies, neither GFAP-nor UB-negative RF were found on immunoelectron microscopy. VIM reaction on IF and a few RF was demonstrated in one tumor processed at low temperature into Lowicryl; it was much weaker than that for GFAP. Many cells with RF contained lysosome-like inclusions with material displaying electron density similar to adjacent RF; few of these inclusions were reactive for UB. It is concluded that RF formation is associated with ubiquitination of astrocytic IF. GFAP-and VIM-immunoreactive IF and products of their disintegration contribute to RF material. It is also suggested that the lysosomal system of astrocytes partially degrades RF.

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References

  1. Bendayan M, Zollinger M (1983) Ultrastructural localization of antigenetic sites on osmium-fixed tissues applying the protein A-gold technique. J Histochem Cytochem 31: 101–109

    Google Scholar 

  2. Bettica AM, Johnson AB (1990) Ultrastructural immunogold labelling of glial filaments in osmicated and unosmicated epoxy-embedded tissue. J Histochem Cytochem 38: 103–109

    Google Scholar 

  3. Borrett D, Becker LE (1985) Alexander's disease. A disease of astrocytes. Brain 108: 367–385

    Google Scholar 

  4. Brett M, Weller RO (1978) Intracellular serum proteins in cerebral gliomas and metastatic tumours. An immunoperoxidase study. Neuropathol Appl Neurobiol 4: 263–270

    Google Scholar 

  5. Ciechanover A, Finley D, Varshavsky A (1984) Ubiquitin dependence of selective protein degradation demonstrated in the mammalian cell cycle mutant s 85. Cell 37: 57–66

    Google Scholar 

  6. Davie JR, Delcuve GP, Nickel BE, Moirier R, Bailey G (1987) Reduced levels of Histones III and IIIb, and unaltered content of methylated DNA in rainbow trout hepatocellular carcinoma chromatin. Cancer Res 47: 5407–5410

    Google Scholar 

  7. Deck JHN, Eng LF, Bigbee J, Woodcock SM (1978) The role of glial fibrillary acidic protein in the diagnosis of central nervous system tumors. Acta Neuropathol (Berl) 42: 183–190

    Google Scholar 

  8. Dinda AK, Sarka C, Roy S (1990) Rosenthal fibers: an immunohistochemical, ultrastructural and immunoelectron microscopic study. Acta Neuropathol 79: 456–460

    Google Scholar 

  9. Doherty FJ, Wassell JA, Mayer RJ (1987) A putative protein sequestration site involving intermediate filaments for protein degradation by autophagy. Studies with microinjected purified glycolytic enzymes in 3T3-L1 cells. Biochem J 241: 793–800

    Google Scholar 

  10. Earl T, Mangiapane H, Billett EE, Mayer JJ (1987) A putative protein-sequestration site involving intermediate filaments for protein degradation by autophagy. Studies with transplanted Sendai-viral envelope proteins in HTC cells. Biochem J 241: 809–815

    Google Scholar 

  11. Finley D, Varshaysky A (1985) The ubiquitin system: functions and mechanisms. Trends Biochem Sci 47: 275–284

    Google Scholar 

  12. French SW, Swierenga HH, Okanoue T, Marceau N (1987) The cytoskeleton of the liver cell in health and disease. In: Farber E, Phillips MJ (eds) Pathogenesis of liver disease, IAP monograph no. 28. Williams and Wilkins, New York, pp 95–112

    Google Scholar 

  13. Gluszcz A, Giernat L, Habryka K, Alwasiak J, Lach B, Papierz (1971) Rosenthal fibers, birefringent gliofibrillary changes and intracellular homogenous conglomerates in tissue cultures of gliomas. Acta Neuropathol (Berl) 17: 54–67

    Google Scholar 

  14. Goldman JE, Corbin E (1988) Isolation of a major protein component of Rosenthal fibers. Am J Pathol 130: 569–578

    Google Scholar 

  15. Goldman JE, Iwaki T, Kume-Iwaki A, Wisniewski T, Liem RKH (1990) Expression of αB-Crystallin in CNS glia. Presented at XI International Congress of Neuropathology, Kyoto, Japan September 2–8, 1990

  16. Gullotta F, Fliedener E (1972) Spongioblastomas, astrocytomas and Rosenthal fibers. Ultrastructural tissue culture and enzyme histochemical investigations. Acta Neuropathol (Berl) 22: 68–78

    Google Scholar 

  17. Hearn SA, Silver MM, Shouldice JA (1985) Immunoelectron microscopic labeling of immunoglobulin in plasma cells after osmium fixation and epoxy embedding. J Histochem Cytochem 33: 1212–1218

    Google Scholar 

  18. Herpers MJHM, Ramackers RCSA, Aldeweireldt J, Moesker O, Slooff J (1986) Co-expression of glial fibrillary acidic protein and vimentin-type intermediate filaments in human astrocytomas. Acta Neuropathol (Berl) 70: 333–339

    Google Scholar 

  19. Hershko A (1983) Ubiquitin: roles in protein modification and breakdown. Cell 34: 11–12

    Google Scholar 

  20. Hershko A, Ciechanover A, Heller H, Haas AL, Rose IA (1980) Proposed role of ATP in protein breakdown; conjugation of proteins with multiple chains of the early peptide of ATP-dependent proteolysis. Proc Natl. Acad Sci USA 77: 1783–1786

    Google Scholar 

  21. Horoupian DS, Kress Y, Yen SH, Gaskin F (1982) Nickel-induced changes and reappraisal of Rosenthal fibers in focal CNS lesions. J Neuropathol Exp Neurol 41: 664–675

    Google Scholar 

  22. Hsu SM, Soban E (1982) Color modification of diaminobenzidine (DAB) precipitation by metallic ions and its application for double immunohistochemistry. J Histochem Cytochem 30: 1079–1082

    Google Scholar 

  23. Hsu SM, Rhine L, Fung H (1981) Use of Avidin-biotin peroxidase complex (ABC) in immunoperoxidase techniques; a comparison between ABC and unlabelled antibody (PAP) procedures. J Histochem Cytochem 29: 577–580

    Google Scholar 

  24. Iwaki T, Kume-Iwaki A, Goldman JE (1989) Tissue distribution of Alfa-B crystallin in rat organs (abstract). J Neuropathol Exp Neurol 48: 363

    Google Scholar 

  25. Janzer RC, Friede RL (1981) Do Rosenthal fibers contain glial fibrillary acidic protein. Acta Neuropathol (Berl) 55: 75–76

    Google Scholar 

  26. Johnson AB, Bettice AM (1986) Rosenthal fibers in Alexander's disease show glial fibrillary acidic protein (GFAP) immunoreactivity with the immunogold staining method (abstract). J Neuropathol Exp Neurol 45: 349

    Google Scholar 

  27. Johnson AB, Bettice AM (1989) On-grid immunogold labelling of glial intermediate filaments in epoxy-embedded tissue. Am J Anat 185: 335–3411

    Google Scholar 

  28. Kellenbergen E, Carlemalm E, Villiger W, Roth J, Garavito RM (1980) Low-denaturation embedding for electron microscopy of thin sections. Chem. Werke Lowi, Waldkraiburg pp 1–59

    Google Scholar 

  29. Kren Y, Gaskin F, Horoupian DS, Brosnan C (1981) Nickel induction of Rosenthal fibers in rat brain. Brain Res 210: 419–425

    Google Scholar 

  30. Kume-Iwaki A, Iwaki T, Liem RKH, Goldman JE (1989) Alfa-B-Crystallin is a major component of Rosenthal Fibers (abstract). J Neuropathol Exp Neurol 48: 377

    Google Scholar 

  31. Lowe J, Blanchard A, Morrell K, Lennox G, Reynolds L, Billett M, Landon M, Mayer RJ (1988) Ubiquitin is a common factor in intermediate filament inclusion bodies of diverse type in men, including those of Parkinson's disease, Pick's disease, and Alzheimer disease, as well as Rosenthal fibers in astrocytomas, cytoplasmic bodies in muscle and Mallory bodies in alcoholic liver disease. J Pathol 155: 9–15

    Google Scholar 

  32. Lowe J, Morell K, Lennox G, Landon M, Mayer RJ (1989) Rosenthal fibres are based on the ubiquitination of glial filaments. Neuropathol Appl Neurobiol 15: 45–53

    Google Scholar 

  33. MacDowell CM (1978) Fixation and processing. In: Trump BJ, Jones RT (eds) Diagnostic electronmicroscopy, vol 1. Wiley Toronto, pp 113–166

    Google Scholar 

  34. Manetto V, Perry G, Tabaton M, Mulvihill P, Fried VA, Smith HT, Gambetti P, Autilio-Gambetti L (1988) Ubiquitin is associated with abnormal cytoplasmic filaments characteristic of neurodegenerative diseases. Proc Natl Acad Sci USA 85: 4501–4505

    Google Scholar 

  35. Manetto V, Abdul-Karim FW, Perry G, Tabaton M, Autilio-Gambetti L, Gambetti P (1989) Selective presence of ubiquitin in intracellular inclusions. Am J Pathol 134: 505–513

    Google Scholar 

  36. McManus JP, Brewer LM (1987) Isolation, localization and peptides of the oncodevelopmental calcium-binding protein, oncomodulin. Methods Enzymol 139: 156–168

    Google Scholar 

  37. Murti KG, Switzer HT, Fried VA (1988) Ubiquitin is a component of the microtubule network. Proc Natl Acad Sci USA 85: 3019–3023

    Google Scholar 

  38. Ohta M, Marceau N, Perry G, Manetto V, Gambetti P, Autilio-Gambetti L, Metuzals J, Kawahara H, Cadrin M, French SW (1988) Ubiquitin is present on cytokeratin intermediate filaments and Mallory bodies of hepatocytes. Lab Invest 59: 848–856

    Google Scholar 

  39. Raimondi AM, Mullan S, Evans GPN (1962) Human brain tumors: an electron microscopic study. J Neurosurg 19: 731–755

    Google Scholar 

  40. Rechsteiner M (1987) Ubiquitin-mediated pathways for intracellular proteolysis. Annu Rev Cell Biol 3: 1–30

    Google Scholar 

  41. Scopsi L, Larsson L (1986) Increased sensitivity in peroxidase immunocytochemistry. A comparative study of a number of peroxidase visualization methods employing a model system. Histochemistry 84: 221–230

    Google Scholar 

  42. Silver MM, Hearn SA (1987) Postembedding immunoelectron microscopy using protein A-gold. Ultrastruct Pathol 11: 693–703

    Google Scholar 

  43. Smith DA, Lantos PL (1985) Immunocytochemistry of cerebellar astrocytomas: with special note on Rosenthal fibers. Acta Neuropathol (Berl) 66: 155–159

    Google Scholar 

  44. Sternberger LA (1979) Immunohistochemistry, 2nd edn. John Wiley and Sons, New York, pp 104–169

    Google Scholar 

  45. Towbin HT, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitorcellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354

    Google Scholar 

  46. Traub P (1985) Intermediate filaments. A review. Springer-Verlag, Berlin, pp 14, 41, 127–129

    Google Scholar 

  47. Van der Meulen JDM, Houthoff HJ, Ebels EJ (1978) Glial fibrillary acidic protein in human gliomas. Neuropathol Appl Neurobiol 4: 177–190

    Google Scholar 

  48. Velasco ME, Dahl D, Roessmann U, Gambetti P (1980) Immunohistochemical localization of glial fibrillary acidic protein in human glial neoplasms. Cancer 45: 484–494

    Google Scholar 

  49. Wilkinson KD, Urban MK, Haas AL (1980) Ubiquitin is the ATP-dependent protolysis factor I of rabbit reticulocytes. J Biochem 225: 7529–7532

    Google Scholar 

  50. Wisniewski T, Iwaki T, Goldman JE (1990) Subcellular localization of α-crystallin B chain in CNS glia (abstract). J Neuropathol Exp Neurol 49: 344

    Google Scholar 

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Authors and Affiliations

  1. Department of Laboratory Medicine (Neuropathology), Ottawa Civic Hospital, K1Y 4E9, Ottawa, Ontario, Canada

    B. Lach, P. Rippstein & A. Gregor

  2. Department of Pathology, University of Ottawa, K1Y 4E9, Ottawa, Ontario, Canada

    B. Lach

  3. Department of Anatomy, University of Ottawa, K1Y 4E9, Ottawa, Ontario, Canada

    W. Staines

  4. Division of Biological Sciences, National Research Council of Canada, Canada

    M. Sikorska

  5. Department of Biochemistry, University of Manitoba, Canada

    T. R. Davie

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  1. B. Lach
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  2. M. Sikorska
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  4. A. Gregor
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  5. W. Staines
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  6. T. R. Davie
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Lach, B., Sikorska, M., Rippstein, P. et al. Immunoelectron microscopy of Rosenthal fibers. Acta Neuropathol 81, 503–509 (1991). https://doi.org/10.1007/BF00310130

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