Summary
Corpora amylacea (CA) accumulation in the central nervous system (CNS) is associated with both normal aging and neurodegenerative conditions such as Alzheimer's disease (AD). CA is reported to be primarily composed of glucose polymers, but approximately 4% of the total weight of CA is consistently composed of protein. CA protein resolved on sodium dodecylsulfatepolyacrylamide gel electrophoresis showed a broad range of polypeptides ranging from 24 to 133 kDa, with four abundant bands. Immunoblots of the profile of polypeptides solubilized from purified CA, showed positive ubiquitin (Ub) immunoreactivity for all the bands. Antisera to heat-shock proteins (hsp) 28 and 70 reacted selectively with bands of 30 and 67 kDa. These results show that Ub is associated with the primary protein components of CA and that the polypeptides are likely to be Ub conjugates. Immunostaining experiments were performed to specifically characterize the protein components of CA in brain tissue sections as well as those of CA purified from both AD and normal aged brains. In all cases CA showed positive reactions with antibodies to Ub, with antibodies raised against either paired helical filaments or hsp 28 or 70, the most prominent staining being with antibodies to Ub, hsp 28 or hsp 70. The presence of Ub and hsp 28 and 70, which are actively induced after stress, suggests that accumulation of altered proteins, possibly attributed to an increased frequency of unusual post-translational modifications or to a sustained physiological stress (related to both normal aging and neurodegenerative process), may be involved in the pathogenesis of CA.
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References
Alder N (1953) On the nature, origin and distribution of the corpora amylacea of the brain with observations on some new staining reactions. J Ment Sci 99:689–697
Ananthan J, Goldberg AL, Voellmy R (1986) Abnormal proteins serve as eukaryotic stress signals and trigger the activation of heat shock genes. Science 232:522–524
Anzil AP, Herrlinger H, Blinzinger K, Kronski D (1974) Intraneuritic corpora amylacea. Virchows Arch [A] 364:297–304
Averback P (1981) Parasynaptic corpora amylacea in the striatum. Arch Pathol Lab Med 105:334–335
Ball E, Karlik CC, Beall CJ, Saville DL, Sparrow JC, Bullard B, Fyrberg EA (1987) Arthrin, a myofibrillar protein of insect flight muscle, is actin-ubiquitin conjugate. Cell 51:221–228
Behrman S, Caroll JD, Janota I, Matthews WB (1969) Progressive supranuclear palsy. Clinico-pathological study of four cases. Brain 92:663–678
Bond U, Schlesinger MJ (1985) Ubiquitin is a heat-shock protein in chicken embryo fibroblasts. Mol Cell Biol 5:949–956
Bond U, Schlesinger MJ (1986) The chicken ubiquitin gene contains a heat-shock promoter and expresses an unstable mRNA in heat shock cells. Mol Cell Biol 6:4602–4610
Busch H, Goldnopf IL (1981) Ubiquitin-protein conjugates. Mol Cell Biochem 40:173–187
Carlson N, Rogers S, Rechsteiner M (1987) Microinjection of ubiquitin: changes in protein degradation in Hela cells subjected to heat shock. J Cell Biol 104:547–555
Cissé S, Lacoste-Royal G, Laperrierre T, Cabana T, Gauvreau D (1991) Ubiquitin is a component of polypeptides purified from corpora amylacea of aged human brain. Neurochem Res 16:429–433
Ellis RS (1920) Norms for some structural changes in the human cerebellum from birth to old age. J Comp Neurol 32:1–3313
Finley D, Varshavsky A (1985) The ubiquitin system: functions and mechanisms. Trends Biochem Sci 10:343–346
Finley D, Ciechanover A, Varshavsky A (1984) Thermolability of ubiquitin-activating enzyme from the mammalian cell cycle mutant ts85. Cell 37:43–55
Fleming PD, Rogers J (1986) Neuropathology of olfactory system in Alzheimer's disease and normal aging. Soc Neurosci Abstr 12:1314
Fleming PD, Cordoza ME, Woods SG, Griesbach EJ, Worcester MA (1987) Corpora amylacea increased in Alzheimer's disease. Neurology [Suppl 1] 37:157
Fried VA, Smith HT, Hildebrandt E, Weiner K (1987) Ubiquitin has intrinsic proteolytic activity: implications for cellular regulation. Proc Natl Acad Sci USA 84:3685–3689
Gambetti P, DiMauro S, Hirt L, Blume RP (1971) Myclonic epilepsy with Lafora bodies: some ultrastructural, histochemical and biochemical aspects. Arch Neurol 25:483–493
Goff SA, Goldberg AL (1985) Production of abnormal proteins in E. coli stimulates transcription of Ion and other heat shock genes. Cell 41:587
Hershko A, Ciechanover A (1986) The Ubiquitin pathway for the degradation of intracellular proteins. Prog Nucleic Acid Res Mol Biol 33:19–56
Hiromi Y, Hotta Y (1985) Actin gene mutations in Drosophila; heatshock activation in the indirect flight muscle. EMBO J 4:1681–1687
Jahnger JH, Haas AL, Ciechanover A, Blonin J, Eisenhauer D, Taylor A (1986) The eye lens has an active ubiquitin-protein conjugation system. J Biol Chem 261:13760–13767
Janeway R, Ravens JR, Pearce LA, Odor DL, Suzuki K (1967) Progressive myoclonus epilepsy with Lafora inclusion bodies. I. Clinical, genetic, histopathologic and biochemical aspects. Arch Neurol 16:565–582
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Liu HM, Burns AC (1985) Mucopolysaccharides in corpora amylacea of normal and diseased brains. J Neuropathol Exp Neurol 44:335
Manetto V, Perry G, Tabaton M, Mulvihill P, Fried V, Smith H, Gambetti P, Aubillio-Gambetti L (1988) Ubiquitin is associated with abnormal cytoplasmic filaments characteristic of neurodegenerative diseases. Proc Natl Acad Sci USA 85:4501–4505
Munro S, Pelham H (1985) What turns on heatshock genes? Nature 317:477–478
Murti KG, Smith HT, Fried VA (1988) Ubiquitin is a component of the microtubule network. Proc Natl Acad Sci USA 85:3019–3023
Palmucci L, Anzil AP, Luh S (1982) Intra-astrocytic glycogen granules and corpora anylacea stain positively for polyglucosans: a cytochemical contribution on the fine structural polymorphism of particulate polysaccharides. Acta Neuropathol (Berl) 57:99–102
Pelham HRB (1984) Hsp 70 accelerates the recovery of nuclear morphology after heat shock. EMBO J 3:3095–3100
Perry G, Mulvihill P, Fried VA, Smith HT, Grundke-Iqbal I, Iqbal K (1989) Immunochemical properties of ubiquitin conjugates in the paired helical filaments of Alzheimer disease. J Neurochem 52:1523–1528
Ramsey HJ (1965) Ultrastructure of copora amylacea. J Neuropathol Exp Neurol 24:25–39
Rechsteiner M (1987) Ubiquitin mediated pathways for intracellular proteolysis. Ann Rev Cell Biol 3:1–30
Robitaille Y, Carpenter S, Karpati G, DiMauro S (1980) A distinct form of adult polyglucosan body disease with massive involvement of central and peripheral neuronal processes and astrocytes. Brain 103:315–336
Roukema PA, Oderkerk CH (1970) Isolation and preliminary characterization of corpora amylacea from human brains. Psychiatr Neurol Neurochir 73:87–89
Sakai M, Austin J, Witmer F, Trueb L (1969) Studies of corpora amylacea. I: Isolation and preliminary characterization by chemical and histochemical techniques. Arch Neurol 21:526–544
Sakai M, Austin J, Witmer F, Trueb L (1969) Corpora amylacea: isolation, characterization and significance. Trans Am Neurol Assoc 94:336–338
Siegelman M, Bond MW, Gallatin WM, St-John T, Smith HT, Fried VA, Weissman IL (1986) Cell surface molecule associated with stimulated homing is a ubiquinated branched-chain glycoprotein. Science 231:823–829
Speiser S, Etlinger JD (1983) ATP proteolysis in reticulocytes extracts by repressing an endogenous inhibitor. Proc Natl Acad Sci USA 80:3577–3580
Stam FC, Roukema PA (1973) Histochemical and biochemical aspects of corpora amylacea. Acta Neuropathol (Berl) 25:95–102
Sternberger LA (1986) Immunocytochemistry, 3rd edn. John Wiley, New York
Steyaert A, Cissé S, Merhi Y, Kalbakji A, Reid N, Gauvreau D, Lacoste-Royal G (1990) Purification and polypeptide composition of corpora amylacea from aged human brain. J Neurosci Methods 31:59–64
St-John T, Gallatin WM, Siegelman M, Smith HT, Fried VA, Weissman IL (1986) Expression cloning of a lymphocyte homing receptor cDNA: ubiquitin is the reactive species. Science 231:845–850
Swerdlow PS, Finley D, Varshavsky A (1986) Enhancement of immunoblot sensitivity by heating of hydrated filters. Anal Biochem 156:147–153
Wang GP, Grundke-Iqbal I, Kascsak RJ, Iqbal K, Wisniewski HM (1984) Alzheimer neurofibrillary tangles: monoclonal antibodies to inherent antigen(s). Acta Neuropathol (Berl) 62:268–275
Yarden Y, Escobedo JA, Kuang WJ, Yang-Feng TL, Daniel TO, Tremble PM, Chen EY, Ando ME, Harkins RN, Francke U, Fried VA, Ullrich A, Williams LT (1986) Structure of the receptor for platelet-derived growth factor helps define a family of closely related growth factor receptors. Nature 323:226–232
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Supported by doctoral fellowships from the Graduate School of Université de Montréal and INRS-Santé and NIH grants AG07552 and AG09282. S. C. is a recipient of these fellowships
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Cisse, S., Perry, G., Lacoste-Royal, G. et al. Immunochemical identification of ubiquitin and heat-shock proteins in corpora amylacea from normal aged and Alzheimer's disease brains. Acta Neuropathol 85, 233–240 (1993). https://doi.org/10.1007/BF00227716
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DOI: https://doi.org/10.1007/BF00227716