title: Single-Molecule Imaging of Individual Amyloid Protein Aggregates in Human Biofluids.
abstract: The misfolding and aggregation of proteins into amyloid fibrils characterizes many neurodegenerative disorders such as Parkinson's and Alzheimer's diseases. We report here a method, termed SAVE (single aggregate visualization by enhancement) imaging, for the ultrasensitive detection of individual amyloid fibrils and oligomers using single-molecule fluorescence microscopy. We demonstrate that this method is able to detect the presence of amyloid aggregates of α-synuclein, tau, and amyloid-β. In addition, we show that aggregates can also be identified in human cerebrospinal fluid (CSF). Significantly, we see a twofold increase in the average aggregate concentration in CSF from Parkinson's disease patients compared to age-matched controls. Taken together, we conclude that this method provides an opportunity to characterize the structural nature of amyloid aggregates in a key biofluid, and therefore has the potential to study disease progression in both animal models and humans to enhance our understanding of neurodegenerative disorders.
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, United Kingdom.
authors.forename: Mathew H
authors.initials: MH
authors.lastname: Horrocks
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, United Kingdom.
authors.forename: Steven F
authors.initials: SF
authors.lastname: Lee
authors.affiliation: Department of Molecular Neuroscience, Institute of Neurology, University College London , Queen Square, London WC1N 3BG, United Kingdom.
authors.forename: Sonia
authors.initials: S
authors.lastname: Gandhi
authors.affiliation: Reta Lila Weston Institute of Neurological Studies, University College London , 1 Wakefield Street, London WC1N 1PJ, United Kingdom.
authors.forename: Nadia K
authors.initials: NK
authors.lastname: Magdalinou
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, United Kingdom.
authors.forename: Serene W
authors.initials: SW
authors.lastname: Chen
authors.affiliation: Division of Brain Sciences, Imperial College of London, Hammersmith Hospital , Du Cane Road, London W12 0NN, United Kingdom.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Devine
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, United Kingdom.
authors.forename: Laura
authors.initials: L
authors.lastname: Tosatto
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, United Kingdom.
authors.forename: Magnus
authors.initials: M
authors.lastname: Kjaergaard
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, United Kingdom.
authors.forename: Joseph S
authors.initials: JS
authors.lastname: Beckwith
authors.affiliation: Department of Molecular Neuroscience, Institute of Neurology, University College London , Queen Square, London WC1N 3BG, United Kingdom.|Clinical Neurochemistry Laboratory, Department of Psychiatry and Neurochemistry, Institute of Neuroscience and Physiology, the Sahlgrenska Academy, University College University of Gothenburg , Mölndal, Sweden.
authors.forename: Henrik
authors.initials: H
authors.lastname: Zetterberg
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, United Kingdom.
authors.forename: Marija
authors.initials: M
authors.lastname: Iljina
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, United Kingdom.
authors.forename: Nunilo
authors.initials: N
authors.lastname: Cremades
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, United Kingdom.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Dobson
authors.affiliation: Department of Molecular Neuroscience, Institute of Neurology, University College London , Queen Square, London WC1N 3BG, United Kingdom.
authors.forename: Nicholas W
authors.initials: NW
authors.lastname: Wood
authors.affiliation: Department of Chemistry, University of Cambridge , Lensfield Road, Cambridge CB2 1EW, United Kingdom.
authors.forename: David
authors.initials: D
authors.lastname: Klenerman
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D058227:Amyloidogenic Proteins / Q000134:cerebrospinal fluid*; D052160:Benzothiazoles; D002942:Circular Dichroism / Q000379:methods; D003952:Diagnostic Imaging / Q000379:methods*; D005260:Female; D006801:Humans; D008297:Male; D018625:Microscopy, Atomic Force / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods*; D008875:Middle Aged; D010300:Parkinson Disease / Q000134:cerebrospinal fluid*; D013844:Thiazoles
chemical_list: D058227:Amyloidogenic Proteins; D052160:Benzothiazoles; D013844:Thiazoles; C009462:thioflavin T
keywords: CSF; Parkinson’s; biomarkers; single-molecule