Detecting Aβ*56 Oligomers in Brain Tissues

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 670)

Abstract

Since its original description in 1906 by Dr Alois Alzheimer, amyloid plaques and neurofibrillary tangles have remained the hypothetical cause of Alzheimer’s disease. However, plaque burden poorly predicts cognitive status in humans, which led several groups to investigate the possibility that soluble species of amyloid-beta (Aβ) peptides could be playing an important pathological function in the aging brain. Through a multistep fractionation protocol, we identified a 56 kDa oligomer of Aβ, termed Aβ*56, the amount of which correlates with cognitive impairment. Here, we describe our biochemical approach to isolate this oligomeric Aβ species in brain tissue of transgenic mouse models of AD.

Key words

Alzheimer Amyloid-beta (Aβ) Brain SDS-PAGE Western blotting Transgenic mouse model 

References

  1. 1.
    Haass, C., and Selkoe, D. J. (2007) Soluble protein oligomers in neurodegeneration: lessons from the Alzheimer’s amyloid beta-peptide. Nat Rev Mol Cell Biol 8, 101–12.PubMedCrossRefGoogle Scholar
  2. 2.
    Walsh, D. M., Minogue, A. M., Sala Frigerio, C., Fadeeva, J. V., Wasco, W., and Selkoe, D. J. (2007) The APP family of proteins: similarities and differences. Biochem Soc Trans 35, 416–20.PubMedCrossRefGoogle Scholar
  3. 3.
    Selkoe, D. J., and Wolfe, M. S. (2007) Presenilin: running with scissors in the membrane. Cell 131, 215–21.PubMedCrossRefGoogle Scholar
  4. 4.
    Lesne, S., Koh, M. T., Kotilinek, L., Kayed, R., Glabe, C. G., Yang, A., Gallagher, M., and Ashe, K. H. (2006) A specific amyloid-beta protein assembly in the brain impairs memory. Nature 440, 352–7.PubMedCrossRefGoogle Scholar
  5. 5.
    Walsh, D. M., and Selkoe, D. J. (2007) A beta oligomers – a decade of discovery. J Neurochem 101, 1172–84.PubMedCrossRefGoogle Scholar
  6. 6.
    Cleary, J. P., Walsh, D. M., Hofmeister, J. J., Shankar, G. M., Kuskowski, M. A., Selkoe, D. J., and Ashe, K. H. (2005) Natural oligomers of the amyloid-beta protein specifically disrupt cognitive function. Nat Neurosci 8, 79–84.PubMedCrossRefGoogle Scholar
  7. 7.
    Walsh, D. M., Klyubin, I., Fadeeva, J. V., Cullen, W. K., Anwyl, R., Wolfe, M. S., Rowan, M. J., and Selkoe, D. J. (2002) Naturally secreted oligomers of amyloid beta protein potently inhibit hippocampal long-term potentiation in vivo. Nature 416, 535–9.PubMedCrossRefGoogle Scholar
  8. 8.
    Shankar, G. M., Li, S., Mehta, T. H., Garcia-Munoz, A., Shepardson, N. E., Smith, I., Brett, F. M., Farrell, M. A., Rowan, M. J., Lemere, C. A., Regan, C. M., Walsh, D. M., Sabatini, B. L., and Selkoe, D. J. (2008) Amyloid-beta protein dimers isolated directly from Alzheimer’s brains impair synaptic plasticity and memory. Nat Med 14, 837–42.PubMedCrossRefGoogle Scholar
  9. 9.
    Cheng, I. H., Scearce-Levie, K., Legleiter, J., Palop, J. J., Gerstein, H., Bien-Ly, N., Puolivali, J., Lesne, S., Ashe, K. H., Muchowski, P. J., and Mucke, L. (2007) Accelerating amyloid-beta fibrillization reduces oligomer levels and functional deficits in Alzheimer disease mouse models. J Biol Chem 282, 23818–28.PubMedCrossRefGoogle Scholar
  10. 10.
    Lesne, S., Kotilinek, L., and Ashe, K. H. (2008) Plaque-bearing mice with reduced levels of oligomeric amyloid-beta assemblies have intact memory function. Neuroscience 151, 745–9.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press 2010

Authors and Affiliations

  1. 1.Department of Neuroscience, Institute for Translational NeuroscienceUniversity of MinnesotaMinneapolisUSA

Personalised recommendations