title: High order W02-reactive stable oligomers of amyloid-β are produced in vivo and in vitro via dialysis and filtration of synthetic amyloid-β monomer.
abstract: Oligomeric forms of amyloid-β (Aβ) are thought to be responsible for the pathogenesis of Alzheimer's disease. While many oligomers of Aβ are thought to be naturally occurring in the brain of humans and/or transgenic animals, it is well known that Aβ oligomers are also readily produced in vitro in the laboratory. In recent studies, we discovered that synthetic monomeric Aβ (4.7 kDa) could be transformed by microdialysis to higher molecular weight species (approximately 56 kDa, by western blot). Surface-enhanced laser desorption/ionization mass spectrometry and electron microscopy further identified these species' as potential Aβ oligomers. The production of similar species could also be produced by centrifugal filtration and this formation was concentration and pore-size dependent. These higher order species of Aβ were resistant to dissolution in NaOH, HFIP, formic acid, urea, and guanidine. We postulate that we have identified a novel way of producing a high order species of oligomeric Aβ and we provide evidence to suggest that Aβ oligomers can quite easily be a product of normal laboratory practices. These data suggest that the experimental detection of higher order oligomers in tissues derived from Alzheimer's disease brains or from animal models of disease could, in some cases, be a product the method of analysis.
authors.affiliation: The Florey Institute of Neuroscience and Mental Health, Division of Mental Health, Parkville, VIC, Australia.
authors.forename: Elysia
authors.initials: E
authors.lastname: Robb
authors.affiliation: The Florey Institute of Neuroscience and Mental Health, Division of Mental Health, Parkville, VIC, Australia The Department of Pharmacology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, VIC, Australia.
authors.forename: Keyla
authors.initials: K
authors.lastname: Perez
authors.affiliation: The Florey Institute of Neuroscience and Mental Health, Division of Mental Health, Parkville, VIC, Australia The Department of Pharmacology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, VIC, Australia.
authors.forename: Lin W
authors.initials: LW
authors.lastname: Hung
authors.affiliation: The Florey Institute of Neuroscience and Mental Health, Division of Mental Health, Parkville, VIC, Australia.
authors.forename: Colin L
authors.initials: CL
authors.lastname: Masters
authors.affiliation: The Florey Institute of Neuroscience and Mental Health, Division of Mental Health, Parkville, VIC, Australia The Department of Pharmacology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, VIC, Australia.
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Barnham
authors.affiliation: The Florey Institute of Neuroscience and Mental Health, Division of Mental Health, Parkville, VIC, Australia.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Cherny
authors.affiliation: The Florey Institute of Neuroscience and Mental Health, Division of Mental Health, Parkville, VIC, Australia.
authors.forename: Ashley I
authors.initials: AI
authors.lastname: Bush
authors.affiliation: The Florey Institute of Neuroscience and Mental Health, Division of Mental Health, Parkville, VIC, Australia.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Adlard
authors.affiliation: The Florey Institute of Neuroscience and Mental Health, Division of Mental Health, Parkville, VIC, Australia.
authors.forename: David I
authors.initials: DI
authors.lastname: Finkelstein
mesh_terms: D000544:Alzheimer Disease / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D016564:Amyloid beta-Protein Precursor / Q000235:genetics; D000818:Animals; D001921:Brain / Q000378:metabolism* / Q000648:ultrastructure; D003956:Dialysis; D004195:Disease Models, Animal; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D046529:Microscopy, Electron, Transmission; D008970:Molecular Weight; D009154:Mutation / Q000235:genetics; D010446:Peptide Fragments / Q000737:chemistry; D053764:Presenilin-1 / Q000235:genetics; D016622:Silver Staining; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D016229:Amyloid beta-Peptides; D016564:Amyloid beta-Protein Precursor; C508853:PSEN1 protein, human; D010446:Peptide Fragments; D053764:Presenilin-1
keywords: Aggregation factors; Alzheimer's disease; W02-reactive; amyloid aggregation; amyloid-beta peptide; filtration; microdialysis; oligomer; resistant