Amyloid β oligomers (AβOs) accumulate early in Alzheimer's disease (AD) and experimentally cause memory dysfunction and the major pathologies associated with AD, for example, tau abnormalities, synapse loss, oxidative damage, and cognitive dysfunction. In order to develop the most effective AβO-targeting diagnostics and therapeutics, the AβO structures contributing to AD-associated toxicity must be elucidated. Here, we investigate the structural properties and pathogenic relevance of AβOs stabilized by the bifunctional crosslinker 1,5-difluoro-2,4-dinitrobenzene (DFDNB). We find that DFDNB stabilizes synthetic Aβ in a soluble oligomeric conformation. With DFDNB, solutions of Aβ that would otherwise convert to large aggregates instead yield solutions of stable AβOs, predominantly in the 50-300 kDa range, that are maintained for at least 12 days at 37°C. Structures were determined by biochemical and native top-down mass spectrometry analyses. Assayed in neuronal cultures and i.c.v.-injected mice, the DFDNB-stabilized AβOs were found to induce tau hyperphosphorylation, inhibit choline acetyltransferase, and provoke neuroinflammation. Most interestingly, DFDNB crosslinking was found to stabilize an AβO conformation particularly potent in inducing memory dysfunction in mice. Taken together, these data support the utility of DFDNB crosslinking as a tool for stabilizing pathogenic AβOs in structure-function studies.
Pubmed ID: 30565253 RIS Download
Publication data is provided by the National Library of Medicine ® and PubMed ®. Data is retrieved from PubMed ® on a weekly schedule. For terms and conditions see the National Library of Medicine Terms and Conditions.
Shared use and training facility. Servies include Confocal Laser Scanning Microscopy, DIC (Differential Interference Contrast Microscopy), FCS (Fluorescence Correlation Spectroscopy), FLIP (Fluorescence-Loss-In-Photobleaching), FRAP (Fluorescence Recovery After Photobleaching),FRET (Fluorescence/ Forster Resonance Energy Transfer),Live-cell Imaging,Phase Contrast Microscopy,Widefield Fluorescence Microscopy,Image Processing and Analysis. Services include poster printing, and specimen preparation. Provides training for most instruments. Instruments include Leica DM6B Fluorescent Microscope (Hogan 5-112), Leica TCS SP8 Confocal Microscope (Hogan 5-128),Leica SP5 II Confocal Microscope (Hogan 5-114),Leica Spinning Disk Confocal Microscope (Hogan 5-113),DeltaVision Deconvolution Microscope (Hogan 5-111),Olympus IX83 Inverted Fluorescent Microscope (Silverman Hall 1-567),Olympus IX53 Inverted Color Microscope (Silverman Hall 1-567), LionHeart Automated Microscope BioTeck (Hogan 5-110).
View all literature mentionsThis unknown targets Tau, phospho (Thr231)
View all literature mentionsThis unknown targets tau, phospho (Ser396)
View all literature mentionsThis monoclonal targets Phospho-Tau (Ser202, Thr205)
View all literature mentionsThis monoclonal targets Na+/K+-ATPase alpha3 (H-4)
View all literature mentionsThis monoclonal secondary targets IgG
View all literature mentionsThis monoclonal targets beta-Amyloid 1-16
View all literature mentionsStatistical analysis software that combines scientific graphing, comprehensive curve fitting (nonlinear regression), understandable statistics, and data organization. Designed for biological research applications in pharmacology, physiology, and other biological fields for data analysis, hypothesis testing, and modeling.
View all literature mentionsMus musculus with name B6SJLF1/J from IMSR.
View all literature mentionsSoftware package as distribution of ImageJ and ImageJ2 together with Java, Java3D and plugins organized into coherent menu structure. Used to assist research in life sciences.
View all literature mentionsSoftware tool for automated microscope acquisition, device control, and image analysis. Used for integrating dissimilar fluorescent microscope hardware and peripherals into a single custom workstation, while providing all the tools needed to perform analysis of acquired images. Offers user friendly application modules for analysis such as cell signaling, cell counting, and protein expression.
View all literature mentionsThis unknown targets tau, phospho (Ser396)
View all literature mentionsSoftware tool for automated microscope acquisition, device control, and image analysis. Used for integrating dissimilar fluorescent microscope hardware and peripherals into a single custom workstation, while providing all the tools needed to perform analysis of acquired images. Offers user friendly application modules for analysis such as cell signaling, cell counting, and protein expression.
View all literature mentionsThis monoclonal secondary targets IgG
View all literature mentions