Amyloid seeds formed by cellular uptake, concentration, and aggregation of the amyloid-beta peptide

Xiaoyan Hu, Scott L. Crick, Guojun Bu, Carl Frieden, Rohit V. Pappu, Jin Moo Lee

Research output: Contribution to journalArticlepeer-review

250 Scopus citations

Abstract

One of the neuropathological hallmarks of Alzheimer's disease (AD) is the amyloid plaque, primarily composed of aggregated amyloidbeta (Aβ) peptide. In vitro, Aβ1-42, the major alloform of Aβ found in plaques, self-assembles into fibrils at micromolar concentrations and acidic pH. Such conditions do not exist in the extracellular fluid of the brain where the pH is neutral and Aβ concentrations are in the nanomolar range. Here, we show that extracellular soluble Aβ (sAβ) at concentrations as low as 1 nM was taken up by murine cortical neurons and neuroblastoma (SHSY5Y) cells but not by human embryonic kidney (HEK293) cells. Following uptake, Aβ accumulated in Lysotracker-positive acidic vesicles (likely late endosomes or lysosomes) where effective concentrations (>2.5 μM) were greater than two orders of magnitude higher than that in the extracellular fluid (25 nM), as quantified by fluorescence intensity using laser scanning confocal microscopy. Furthermore, SHSY5Y cells incubated with 1βM Aβ1-42 for several days demonstrated a time-dependent increase in intracellular high molecular weight (HMW) (>200 kDa) aggregates, which were absent in cells grown in the presence of Aβ1-40. Homogenates from these Aβ1-42-loaded cells were capable of seeding amyloid fibril growth. These results demonstrate that Aβ can be taken up by certain cells at low physiologically relevant concentrations of extracellular Aβ, and then concentrated into endosomes/lysosomes. At high concentrations, vesicular Aβ aggregates to form HMW species which are capable of seeding amyloid fibril growth.We speculate that extrusion of these aggregates may seed extracellular amyloid plaque formation during AD pathogenesis.

Original languageEnglish (US)
Pages (from-to)20324-20329
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume106
Issue number48
DOIs
StatePublished - Dec 1 2009

Keywords

  • Amyloid fibrils
  • Late endosomes
  • Lysosomes
  • Plaques

ASJC Scopus subject areas

  • General

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