Endoplasmic reticulum and trans-Golgi network generate distinct populations of Alzheimer β-amyloid peptides

Research output: Contribution to journalArticle

Abstract

The excessive generation and accumulation of 40- and 42-aa β-amyloid peptides (Aβ40/Aβ42) in selectively vulnerable brain regions is a major neuropathological feature of Alzheimer's disease. Aβ, derived by proteolytic cleavage from the β-amyloid precursor protein (βAPP), is normally secreted. However, recent evidence suggests that significant levels of Aβ also may remain inside cells. Here, we have investigated the subcellular compartments within which distinct amyloid species are generated and the compartments from which they are secreted. Three experimental approaches were used: (i) immunofluorescence performed in intact cortical neurons; (ii) sucrose gradient fractionation performed with mouse neuroblastoma cells stably expressing wild-type βAPP695 (N2a695); and (iii) cell-free reconstitution of Aβ generation and trafficking from N2a695 cells. These studies demonstrate that: (i) Aβ40 (Aβ1-40 plus Aβ(x-40), where x is an NH2-terminal truncation) is generated exclusively within the trans-Golgi Network (TGN) and packaged into post-TGN secretory vesicles; (ii) Aβ(x-42) is made and retained within the endoplasmic reticulum in an insoluble state; (iii) Aβ42 (Aβ1-42 plus Aβ(x-42)) is made in the TGN and packaged into secretory vesicles; and (iv) the amyloid peptides formed in the TGN consist of two pools (a soluble population extractable with detergents and a detergent-insoluble form). The identification of the organelles in which distinct forms of Aβ are generated and from which they are secreted should facilitate the identification of the proteolytic enzymes responsible for their formation.

Details

Authors
  • Jeffrey P. Greenfield
  • Julia Tsai
  • Gunnar K. Gouras
  • Bing Hai
  • Gopal Thinakaran
  • Frederic Checler
  • Sangram S. Sisodia
  • Paul Greengard
  • Huaxi Xu
External organisations
  • Rockefeller University
  • Weill Cornell Medical College
  • University of Chicago
  • CNRS Institute de Pharmacologie Moleculaire et Cellulaire
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Neurosciences
Original languageEnglish
Pages (from-to)742-747
JournalProceedings of the National Academy of Sciences of the United States of America
Volume96
Issue number2
Publication statusPublished - 1999 Jan 19
Publication categoryResearch
Peer-reviewedYes
Externally publishedYes