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Modifications of Phosphorylated Tau Immunoreactivity Linked to Excitotoxicity in Neuronal Cultures

  • J. Hugon
  • P. Sindou
  • M. Lesort
  • P. Couratier
  • F. Esclaire
  • C. Yardin
Conference paper
Part of the Research and Perspectives in Alzheimer’s Disease book series (ALZHEIMER)

Abstract

Glutamate is one of the major excitatory neurotransmitters in the human brain (Fonnum 1984) but is also a potent neurotoxin producing in vitro and in vivo neuronal degradation and death (Meldrum and Garthwaite 1991). Three types of post-synaptic receptors are described: N-Methyl D aspartate, AMPA/Kainate, and metabotropic according to their principal pharmacological agonists. A large variety of molecules can activate these three post-synaptic receptors (Seeburg 1993). Glutamate has been implicated in the pathophysiology of many neurodegenerative disorders both acute, such as stroke and hypoglycemia (Simon et al. 1984; Wieloch 1985), and chronic, such as amyotrophic lateral sclerosis (Couratier et al. 1993), Parkinson’s disease (Turski et al. 1991), Huntington’s disease (Young et al. 1988), AIDS dementia complex (Lipton et al. 1991) and Alzheimer’s disease (Koh et al. 1990; Kowall and Beal 1991; Mattson et al. 1992). Glutamate is also able to produce an intracellular signal transduction into neurons leading to protein phosphorylations (Scholz and Palfrey 1991). Tau is a microtubule-associated protein which favours microtubule polymerisation and stabilisation (Kosik 1993). One of the major neuropathological hallmarks of Alzheimer’s disease is neurofibrillary tangles (NFT) associated with neuronal degeneration. NFT are composed of paired helical neurofilaments (PHF). Tau is one of the principal constituents of PHF but PHF tau is abnormally phosphorylated (Brion et al. 1991; Goedert 1993; Lee and Trojanowski 1992). The goal of the following experiments was to detect changes in tau immunoreactivity observed in neuronal cultures after glutamate exposure. We used a new monoclonal antibody, AT8 (provided by Innogenetics), raised against an abnormally phophorylated tau site at serine 202. Immunocytochemistry with confocal laser microscopy and immunoblot studies were carried out in these experiments.

Keywords

Amyotrophic Lateral Sclerosis Neuronal Culture Paired Helical Filament Glutamate Toxicity Glutamate Exposure 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Brion JP, Hanger DP, Couck AM, Anderton BH (1991) A68 proteins in Alzheimer’s disease are composed of several tau isoforms in a phosphorylated state which affects their electrophoretic mobilities. Biochem J 279: 831–836PubMedGoogle Scholar
  2. Choi DW (1987) Ionic dependence of glutamate neurotoxicity. J Neurosci 7: 369–379PubMedGoogle Scholar
  3. Couratier P, Hugon J, Sindou P, Vallat JM, Dumas M (1993) Cell culture evidence for neuronal degeneration in amyotrophic lateral sclerosis being linked to glutamate AMPA/ Kainate receptors. Lancet 341: 265–268PubMedCrossRefGoogle Scholar
  4. Delacourte A, Defossez A (1986) Alzheimer’s disease: tau proteins, the promoting factors of microtubule assembly, are major components of paired helical filaments. J Neurol Sci 76: 173–186PubMedCrossRefGoogle Scholar
  5. Fonnum F (1984) Glutamate: a neurotransmitter in mammalian brain. J Neurochem 42: 1–11PubMedCrossRefGoogle Scholar
  6. Goedert M (1993) Tau protein and the neurofibrillary pathology of Alzheimer’s disease. Trends Neurosci 16: 460–465PubMedCrossRefGoogle Scholar
  7. Goedert M, Jakes R, Crowther RA, Six J, Lobke U, Vandermeeren I, Cras P, Troganows JQ, Lee VM-Y (1993) The abnormal phosphorylation of tau protein at Ser-202 in Alzheimer disease recapitulates phosphorylation during development. Proc Natl Acad Soc USA 90: 5066–5070CrossRefGoogle Scholar
  8. Kenessey A, Yen S-H C (1993) The extent of phosphorylation of fetal tau is comparable to that of PHF-tau from Alzheimer paired helical filaments. Brain Res 629: 40–46PubMedCrossRefGoogle Scholar
  9. Koh JY, Yang LL, Cotman CW (1990) ß-amyloid protein increases the vulnerability of cultured cortical neurons to excitotoxic damage. Brain Res 533: 315–320PubMedCrossRefGoogle Scholar
  10. Kosik KS (1993) The molecular and cellular biology of Tau. Brain Pathol 3: 39–43PubMedCrossRefGoogle Scholar
  11. Kowall NW, Beal MF (1991) Glutamate, glutaminase and taurine immunoreactive neurons develop neurofibrillary tangles in Alzheimer’s disease. Ann Neurol 29: 162–167PubMedCrossRefGoogle Scholar
  12. Lee V M-Y, Trojanowski JO (1992) The disordered neuronal cytoskeleton in Alzheimer’s disease. Curr Opin Neurobiol 2: 653–656PubMedCrossRefGoogle Scholar
  13. Lipton SA, Sucher NJ, Kaiser PK, Dreyer EB (1991) Synergistic effects of HIV coat protein and NMDA receptor-mediated neurotoxicity. Neuron 7: 111–118PubMedCrossRefGoogle Scholar
  14. Mattson MP (1990) Antigenic changes similar to those seen in neurofibrillary tangles are elicited by glutamate and Ca2+ influx in cultured hippocampal neurons. Neuron 2: 105–117CrossRefGoogle Scholar
  15. Mattson MP, Cheng B, Davis D, Bryant K, Lieberbury I, Rydel A (1992) ß-amyloid peptides destabilize calcium homeostasis and render human cortical neurons vulnerable to exocitotoxicity. J Neurosci 12: 376–389PubMedGoogle Scholar
  16. Meldrum B, Garthwaite J (1991) Excitatory amino acid neurotoxicity and neurodegenerative disease. TIPS Spec Rept 54–62Google Scholar
  17. Sautiere PE, Sindou P, Couratier P, Hugon J, Wattez A, Delacourte A (1992) Tau antigenic changes induced by glutamate in rat primary culture model: a biochemical approach. Neurosci Lett 140: 206–210PubMedCrossRefGoogle Scholar
  18. Scholz WK, Palfrey HC (1991) Glutamate-stimulated protein phosphorylation in cultured hippocampal pyramidal neurons. J Neuronsci 11: 2422–2432Google Scholar
  19. Seeburg PH (1993) The molecular biology of mammalian glutamate receptor channels. TINS 16: 359–365PubMedGoogle Scholar
  20. Simon RP, Swan JH, Griffiths T, Meldrum BS (1984) Blockade of N-methyl-D-aspartate receptors may protect against ischemic damage in the brain. Science 2267: 850–852CrossRefGoogle Scholar
  21. Sindou P, Couratier P, Barthe D, Hugon J (1992) A dose-dependent increase of tau immunostaining is produced by glutamate toxicity in primary neuronal cultures. Brain Res 572: 242–246PubMedCrossRefGoogle Scholar
  22. Turski L, Bressler K, Rettig K-J, Loeschmann PA, Wachtel H (1991) Protection of substantia nigra from MPP+ neurotoxicity by N-methyl-D-aspartate antagonists. Nature 349: 414–419PubMedCrossRefGoogle Scholar
  23. Wieloch T (1985) Hypoglycemia-induced neuronal damage prevented by an N-methyl-D-aspartate antagonist. Science 230: 681–683PubMedCrossRefGoogle Scholar
  24. Young AB, Greenamyre JT, Hollingworth Z, Albin R, d’Amato C, Shoulson I, Penney JQ (1988) NMDA receptor losses in putamen from patients with Huntington’s disease. Science 241: 981–983PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1995

Authors and Affiliations

  • J. Hugon
    • 1
  • P. Sindou
  • M. Lesort
  • P. Couratier
  • F. Esclaire
  • C. Yardin
  1. 1.Unité de Neurobiologie Cellulaire, Laboratoire d’HistologieFaculté de MédecineLimoges CédexFrance

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