Acta Neuropathologica

, Volume 75, Issue 4, pp 345–353 | Cite as

Lewy bodies are ubiquitinated

A light and electron microscopic immunocytochemical study
  • S. Kuzuhara
  • H. Mori
  • N. Izumiyama
  • M. Yoshimura
  • Y. Ihara
Regular Papers


The nature of Lewy bodies (LBs) in the brain stem and cerebral cortex in five cases of diffuse Lewy body disease and one case of Parkinson's disease with dementia were investigated immunocytochemically with various antibodies to cytoskeletal proteins, paired helical filaments (PHF) and ubiquitin. Antibodies to 200-kDa component of neurofilament, tau and PHF showed no significant reactions with most of LBs. Antibodies to high-molecular weight microtubule-associated proteins (HMWMAPs) moderately stained the periphery of a few of LBs. A monoclonal antibody to PHF (DF2) which recognizes ubiquitin, and polyclonal antibodies to ubiquitin immunostained virtually all of the typical and cortical LBs as intensely as Alzheimer's neurofibrillary tangles and senile plaque neurites: the periphery of LBs was darkly stained, whereas the central core of typical LBs and central zone of cortical LBs were less intensely stained or remained unstained. Immunoelectron microscopy of the LBs with DF2 revealed that immune reaction products were located on the filaments exclusively in the periphery of LBs, but not on those in the center. These findings suggest that both types of LBs are immunocytochemically indistinguishable despite some structural differences, and that peripherally located filaments in LBs are tagged with ubiquitin, an element required for the ATP-dependent proteolysis system in the cell. Antibodies to ubiquitin are the most useful marker of LBs ever known.

Key words

Lewy body Immunocytochemistry Ubiquitin Paired helical filaments 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Ciechanover A, Finley D, Varshavsky A (1984) Ubiquitin dependence of selective protein degradation demonstrated in the mammalian cell cycle mutant ts85. Cell 37:57–66Google Scholar
  2. 2.
    Dickson DW, Kress V, Crowe A, Yen S-H (1985) Monoclonal antibodies to Alzheimer neurofibrillary tangles. 2. Demonstration of common antigenic determinant between ANT and neurofibrillary degeneration in progressive supranuclear palsy. Am J Pathol 120:292–303Google Scholar
  3. 3.
    Duffy PE, Tennyson VM (1965) Phase and electron microscopic observation of Lewy bodies and melanin granules in the substantia nigra and locus caeruleus in Parkinson's disease. J Neuropathol Exp Neurol 24:398–414Google Scholar
  4. 4.
    Forno LS (1986) The Lewy body in Parkinson's disease Adv Neurol 45:35–43Google Scholar
  5. 5.
    Forno LS, Langston JW, Delanney LE, Irwin I, Ricaurte GA (1986a) Locus ceruleus lesions and eosinophilic inclusions in MPTP-treated monkeys Ann Neurol 20:449–455Google Scholar
  6. 6.
    Forno LS, Sternberger LA, Sternberger NH, Strefling AM, Swanson K, Eng LF (1986b) Reaction of Lewy bodies with antibodies to phosphorylated and nonphosphorylated neurofilaments. Neurosci Lett 64:253–258Google Scholar
  7. 7.
    Gambetti P, Autilio-Gambetti L, Papasozomenos SC (1981) Bodian's silver method stains neurofilament polypeptides. Science 213:1521–1522Google Scholar
  8. 8.
    Goldman JE, Yen S-H (1983) Lewy bodies of Parkinson's disease contain neurofilament antigens. Science 221:1082–1084Google Scholar
  9. 9.
    Grundke-Iqbal I, Iqbal K, Quinlan M, Tung Y-C, Zaidi MS, Wisniewski HM (1986a) Microtubule-associated protein tau, a component of Alzheimer paired helical filaments. J Biol Chem 261:6084–6089Google Scholar
  10. 10.
    Grundke-Iqbal I, Iqbal K, Tung Y-C, Quinlan M, Wisniewski HM, Binder LI (1986b) Abnormal phosphorylation of the microtubule-associated protein tau in Alzheimer cytoskeletal pathology. Proc Natl Acad Sci USA 83:4913–4917Google Scholar
  11. 11.
    Haas AL, Bright PM (1985) The immunochemical detection and quantitation of intracellular ubiquitin-protein conjugates. J Biol Chem 260:12464–12473Google Scholar
  12. 12.
    Hershko A (1983) Ubiquitin: roles in protein modification and breakdown. Cell 34:11–12Google Scholar
  13. 13.
    Hershko A, Eytan E, Ciechanover A, Haas AL (1982) Immunochemical analysis of the turnover of ubiquitin-protein conjugates in intact cells. J Biol Chem 257:13964–13970Google Scholar
  14. 14.
    Hsu S-M, Raine L, Fanger H (1981) Use of avidin-biotinperoxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem 29:577–580Google Scholar
  15. 15.
    Ihara Y, Nukina N, Sugita H, Toyokura Y (1981) Staining of Alzheimer's neurofibrillary tangles with antiserum against 200K component of neurofilament. Proc Jpn Acad [B]57:152–156Google Scholar
  16. 16.
    Ihara Y, Abraham C, Selkoe DJ (1983) Antibodies to paired helical filaments in Alzheimer's disease do not recognize normal brain proteins. Nature 304:727–730Google Scholar
  17. 17.
    Ihara Y, Nukina N, Miura R, Ogawara M (1986) Phosphorylated tau protein is integrated into paired helical filaments in Alzheimer's disease. J Biochem 99:1807–1810Google Scholar
  18. 18.
    Kosaka K (1978) Lewy bodies in cerebral cortex. Report of three cases. Acta Neuropathol (Berl) 42:127–134Google Scholar
  19. 19.
    Kosaka K, Yoshimura M, Ikeda K, Budka H (1984) Diffuse type of Lewy body disease: progressive dementia with abundant cortical Lewy bodies and senile changes of varying degree. A new disease? Clin Neuropathol 3:185–192Google Scholar
  20. 20.
    Langston JW, Ballard P (1983) Chronic parkinsonism in humans due to a product of meperidine-analog synthesis. Science 219:979–980Google Scholar
  21. 21.
    Matus A, Bernhardt R, Jones TH (1981) High-molecular weight microtubule-associated proteins are preferentially associated with dendritic microtubules in brain. Proc Natl Acad Sci USA 78:3010–3014Google Scholar
  22. 22.
    Mori H, Kondo J, Ihara Y (1987) Ubiquitin is a component of paired helical filaments in Alzheimer's disease. Science 235:1641–1644Google Scholar
  23. 23.
    Nukina N, Ihara Y (1983a) Immunocytochemical study on senile plaques in Alzheimer's disease. I. Preparation of an anti-microtubule-associated proteins (MAPs) antiserum and its specificity. Proc Jpn Acad [B] 59:284–287Google Scholar
  24. 24.
    Nukina N, Ihara Y (1983b) Immunocytochemical study on senile plaques in Alzheimer's disease. II. Abnormal dendrites in senile plaques as revealed by anti-microtubule-associated proteins (MAPs) immunostaining. Proc Jpn Acad [B] 59:288–292Google Scholar
  25. 25.
    Nukina N, Ihara Y (1986) One of the antigenic determinants of paired helical filaments is related to tau protein. J Biochem (Tokyo) 99:1541–1544Google Scholar
  26. 26.
    Snyder SH, D'Amato RJ (1986) MPTP: a neurotoxin relevant to the pathophysiology of Parkinson's disease. The 1985 George C. Cotzias lecture. Neurology 36:250–258Google Scholar
  27. 27.
    Sternberger NH, Sternberger LA (1983) Monoclonal antibodies distinguish phosphorylated and nonphosphorylated forms of neurofilaments in situ. Proc Natl Acad Sci USA 80:6126–6130Google Scholar
  28. 28.
    Yoshimura M (1983) Cortical changes in the parkinsonian brain: a contribution to the delineation of “diffuse Lewy body disease”. J Neurol 229:17–32Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • S. Kuzuhara
    • 1
    • 3
  • H. Mori
    • 2
  • N. Izumiyama
    • 3
  • M. Yoshimura
    • 3
  • Y. Ihara
    • 2
  1. 1.Department of NeurologyTokyo Metropolitan Geriatric HospitalItabashi-ku, TokyoJapan
  2. 2.Department of Clinical PhysiologyTokyo Metropolitan Institute of GerontologyItabashi-ku, TokyoJapan
  3. 3.Department of Clinical PathologyTokyo Metropolitan Institute of GerontologyItabashi-ku, TokyoJapan

Personalised recommendations