title: An improved method for generating consistent soluble amyloid-beta oligomer preparations for in vitro neurotoxicity studies.
abstract: Soluble Abeta oligomers are recognized as playing a key role in Alzheimer's disease (AD) pathophysiology. Despite their significance, many investigators encounter difficulty generating reliable preparations for in vitro and in vivo experiments. Solutions of Abeta are often unstable and soluble conformer profiles inconsistent. In this study we describe detailed methods for preparing Abeta oligomers that are stable for several weeks and are enriched for low and high molecular weight oligomeric forms, including the 56-kDa form, a conformer implicated in AD-related cognitive impairment. We characterize their structural and functional properties using Western blot, dot blot, atomic force microscopy, Thioflavine T fluorescence, and primary neuronal culture toxicity assays. These synthetic preparations should prove valuable to many studying Abeta-mediated mechanisms underlying AD.
authors.affiliation: Interdepartmental Graduate Program in Neuroscience, Rochester, NY 14642, USA.
authors.forename: Deborah A
authors.initials: DA
authors.lastname: Ryan
authors.forename: Wade C
authors.initials: WC
authors.lastname: Narrow
authors.forename: Howard J
authors.initials: HJ
authors.lastname: Federoff
authors.forename: William J
authors.initials: WJ
authors.lastname: Bowers
mesh_terms: D016229:Amyloid beta-Peptides / Q000138:chemical synthesis* / Q000378:metabolism / Q000648:ultrastructure; D000818:Animals; D015153:Blotting, Western; D016923:Cell Death; D045744:Cell Line, Tumor; D002470:Cell Survival; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005453:Fluorescence; D015151:Immunoblotting; D051379:Mice; D018625:Microscopy, Atomic Force; D008970:Molecular Weight; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D055503:Protein Multimerization; D012995:Solubility; D013997:Time Factors
chemical_list: D016229:Amyloid beta-Peptides
references.citation: J Neurosci. 2002 Mar 1;22(5):1858-67
references.pmid: 11880515
references.citation: J Neurochem. 2008 Jan;104(2):457-68
references.pmid: 17953662
references.citation: Nat Neurosci. 2002 May;5(5):452-7
references.pmid: 11941374
references.citation: Nat Med. 1995 Oct;1(10):1062-6
references.pmid: 7489364
references.citation: Nat Med. 2008 Aug;14(8):837-42
references.pmid: 18568035
references.citation: Proc Natl Acad Sci U S A. 2004 Nov 16;101(46):16180-5
references.pmid: 15534217
references.citation: J Mol Biol. 2006 Apr 21;358(1):106-19
references.pmid: 16499926
references.citation: J Neurosci. 2002 Aug 1;22(15):6331-5
references.pmid: 12151510
references.citation: J Biol Chem. 1997 Aug 29;272(35):22364-72
references.pmid: 9268388
references.citation: Biochemistry. 1992 Jun 23;31(24):5621-31
references.pmid: 1610809
references.citation: Mol Neurodegener. 2009 Nov 23;4:48
references.pmid: 19930651
references.citation: J Neurochem. 2007 Jan;100(1):23-35
references.pmid: 17116235
references.citation: Proc Natl Acad Sci U S A. 2003 Jan 7;100(1):330-5
references.pmid: 12506200
references.citation: Neurobiol Dis. 2008 May;30(2):212-20
references.pmid: 18353662
references.citation: Proc Natl Acad Sci U S A. 1998 May 26;95(11):6448-53
references.pmid: 9600986
references.citation: J Biol Chem. 2003 Mar 28;278(13):11612-22
references.pmid: 12499373
references.citation: J Biol Chem. 1999 Sep 3;274(36):25945-52
references.pmid: 10464339
references.citation: Mol Neurodegener. 2007 Sep 26;2:18
references.pmid: 17897471
references.citation: Arch Neurol. 2003 Aug;60(8):1119-22
references.pmid: 12925369
references.citation: Science. 2001 Aug 24;293(5534):1449-54
references.pmid: 11520976
references.citation: Protein Sci. 1993 Mar;2(3):404-10
references.pmid: 8453378
references.citation: J Biol Chem. 2001 Sep 14;276(37):35176-84
references.pmid: 11441003
references.citation: Nature. 2006 Mar 16;440(7082):352-7
references.pmid: 16541076
references.citation: Nat Neurosci. 2005 Jan;8(1):79-84
references.pmid: 15608634
references.citation: Nat Med. 1996 Aug;2(8):864-70
references.pmid: 8705854
references.citation: Nat Med. 2005 May;11(5):556-61
references.pmid: 15834427
references.citation: J Biol Chem. 2007 Apr 6;282(14):10311-24
references.pmid: 17284452
references.citation: Am J Pathol. 1999 Sep;155(3):853-62
references.pmid: 10487842
references.citation: J Neurochem. 2004 Jul;90(1):62-9
references.pmid: 15198667
references.citation: Cell Tissue Res. 1997 Apr;288(1):119-26
references.pmid: 9042779
references.citation: Science. 2003 Apr 18;300(5618):486-9
references.pmid: 12702875
references.citation: Proc Natl Acad Sci U S A. 2003 Sep 2;100(18):10417-22
references.pmid: 12925731
references.citation: J Neurosci. 2004 Nov 10;24(45):10191-200
references.pmid: 15537891
references.citation: Neurobiol Aging. 2006 Oct;27(10):1372-84
references.pmid: 16289476
references.citation: J Neurochem. 2001 Nov;79(3):595-605
references.pmid: 11701763
references.citation: J Neurochem. 2005 Nov;95(3):834-47
references.pmid: 16135089