title: Proteomic analysis of the hippocampus in Alzheimer's disease model mice by using two-dimensional fluorescence difference in gel electrophoresis.
abstract: We previously identified the E693Δ mutation in amyloid precursor protein (APP) in patients with Alzheimer's disease (AD) and then generated APP-transgenic mice expressing this mutation. As these mice possessed abundant Aβ oligomers from 8 months of age but no amyloid plaques even at 24 months of age, they are a good model to study pathological effects of amyloid β (Aβ) oligomers. The two-dimensional fluorescence difference in gel electrophoresis (2D-DIGE) technology, using a mixed-sample internal standard, is now recognized as an accurate method to determine and quantify proteins. In this study, we examined the proteins for which levels were altered in the hippocampus of 12-month-old APP(E693Δ)-transgenic mice using 2D-DIGE and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Fourteen proteins were significantly changed in the hippocampus of APP(E693Δ)-transgenic mice. Actin cytoplasmic 1 (β-actin), heat shock cognate 71kDa, γ-enolase, ATP synthase subunit β, tubulin β-2A chain, clathrin light chain B (clathrin) and dynamin-1 were increased. Heat shock-related 70kDa protein 2, neurofilament light polypeptide (NFL), stress-induced-phosphoprotein 2, 60kDa heat shock protein (HSP60), α-internexin, protein kinase C and casein kinase substrate in neurons protein 1 (Pacsin 1), α-enolase and β-actin were decreased. Western blotting also validated the changed levels of HSP60, NFL, clathrin and Pacsin 1 in APP(E693Δ)-transgenic mice. The identified proteins could be classified as cytoskeleton, chaperons, neurotransmission, energy supply and signal transduction. Thus, proteomics by 2D-DIGE and LC-MS/MS has provided knowledge of the levels of proteins in the early stages of AD brain.
authors.affiliation: Laboratory of Molecular Cellular Biology, School of Pharmaceutical Sciences, Kobe Gakuin University, 1-1-3 Minatojima, Chuo-ku, Kobe 650-8586, Japan.
authors.forename: Masaoki
authors.initials: M
authors.lastname: Takano
authors.forename: Takuya
authors.initials: T
authors.lastname: Yamashita
authors.forename: Kazuya
authors.initials: K
authors.lastname: Nagano
authors.forename: Mieko
authors.initials: M
authors.lastname: Otani
authors.forename: Kouji
authors.initials: K
authors.lastname: Maekura
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Kamada
authors.forename: Shin-Ichi
authors.initials: S
authors.lastname: Tsunoda
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Tsutsumi
authors.forename: Takami
authors.initials: T
authors.lastname: Tomiyama
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Mori
authors.forename: Kenji
authors.initials: K
authors.lastname: Matsuura
authors.forename: Shogo
authors.initials: S
authors.lastname: Matsuyama
mesh_terms: D000544:Alzheimer Disease / Q000378:metabolism*; D016564:Amyloid beta-Protein Precursor / Q000235:genetics; D000818:Animals; D002853:Chromatography, Liquid; D005453:Fluorescence; D006624:Hippocampus / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D020543:Proteome / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D057825:Two-Dimensional Difference Gel Electrophoresis
chemical_list: D016564:Amyloid beta-Protein Precursor; D020543:Proteome