Disruption of amyloid plaques integrity affects the soluble oligomers content from Alzheimer disease brains

PLoS One. 2014 Dec 8;9(12):e114041. doi: 10.1371/journal.pone.0114041. eCollection 2014.

Abstract

The implication of soluble Abeta in the Alzheimer's disease (AD) pathology is currently accepted. In fact, the content of soluble extracellular Abeta species, such as monomeric and/or oligomeric Abeta, seems to correlate with the clinico-pathological dysfunction observed in AD patients. However, the nature (monomeric, dimeric or other oligomers), the relative abundance, and the origin (extra-/intraneuronal or plaque-associated), of these soluble species are actually under debate. In this work we have characterized the soluble (defined as soluble in Tris-buffered saline after ultracentrifugation) Abeta, obtained from hippocampal samples of Braak II, Braak III-IV and Braak V-VI patients. Although the content of both Abeta40 and Abeta42 peptides displayed significant increase with pathology progression, our results demonstrated the presence of low, pg/µg protein, amount of both peptides. This low content could explain the absence (or below detection limits) of soluble Abeta peptides detected by western blots or by immunoprecipitation-western blot analysis. These data were in clear contrast to those published recently by different groups. Aiming to explain the reasons that determine these substantial differences, we also investigated whether the initial homogenization could mobilize Abeta from plaques, using 12-month-old PS1xAPP cortical samples. Our data demonstrated that manual homogenization (using Dounce) preserved the integrity of Abeta plaques whereas strong homogenization procedures (such as sonication) produced a vast redistribution of the Abeta species in all soluble and insoluble fractions. This artifact could explain the dissimilar and somehow controversial data between different groups analyzing human AD samples.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Aged, 80 and over
  • Alzheimer Disease / metabolism*
  • Alzheimer Disease / pathology*
  • Amyloid beta-Peptides / chemistry
  • Amyloid beta-Peptides / metabolism*
  • Autopsy
  • Brain / metabolism*
  • Brain / pathology*
  • Cell Line
  • Female
  • Hippocampus / metabolism
  • Hippocampus / pathology
  • Humans
  • Male
  • Plaque, Amyloid*
  • Protein Aggregation, Pathological*

Substances

  • Amyloid beta-Peptides

Grants and funding

This work was supported by Fondo de Investigación Sanitaria (FIS), from Instituto de Salud Carlos III of Spain, through grants PI12/01431 (to AG) and PI12/01439 (to JV), by Junta de Andalucia (collaborative grant CTS-2035 to JV and AG), and by Centro de Investigacion Biomedica en Red Sobre Enfermedades Neurodegenerativas (CIBERNED) (collaborative grants PI2010/08 and PI2013/01 to JV and AG). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.