Intracellular Aβ is increased by okadaic acid exposure in transfected neuronal and non-neuronal cell lines

Xiaoyan Sun, Gregory M. Cole, Teresa Chu, Weiming Xia, Douglas Galasko, Haruyasu Yamaguchi, Kentaro Tanemura, Sally A. Frautschy, Akihiko Takashima

Research output: Contribution to journalArticle

30 Scopus citations

Abstract

Intracellular Aβ was examined in both a neuronal cell line (B103) expressing human APP with Swedish mutation and a non-neuronal cell line (Chinese hamster ovary, CHO) expressing wild human APP. Exposure of the APP695sw-transfected B103 cells to okadaic acid for 3 h, Aβ immunostaining was enhanced, as demonstrated by two independent anti-Aβ antibodies. The confocal microscopic study revealed that the immunoreactivity of Aβ was mainly colocalized with a Golgi marker and partially with an ER marker. Quantitative analyses, using Aβ sandwich ELISA, showed significantly increased intracellular Aβ. False positive detection of Aβ by antibody cross-reaction with APP was ruled out by extracting the fraction with formic acid and making it alkaline before subjecting it to ELISA. This procedure resulted in a fraction that contained little APP. Using CHO cells, OA treatment was also shown to be effective in increasing Aβ, as demonstrated by Western blot. The increased full-length APP and decreased APPC99 were also observed. This is the first study to demonstrate that OA treatment significantly increases intracellular Aβ.

Original languageEnglish (US)
Pages (from-to)195-203
Number of pages9
JournalNeurobiology of Aging
Volume23
Issue number2
DOIs
StatePublished - 2002

Keywords

  • Alzheimer's disease
  • Amyloid
  • Intracellular Aβ
  • Okadaic acid

ASJC Scopus subject areas

  • Clinical Neurology
  • Biological Psychiatry
  • Developmental Neuroscience
  • Neurology
  • Psychology(all)

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