Single-Molecule Imaging of Individual Amyloid Protein Aggregates in Human Biofluids

ACS Chem Neurosci. 2016 Mar 16;7(3):399-406. doi: 10.1021/acschemneuro.5b00324. Epub 2016 Feb 4.

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.

Keywords: CSF; Parkinson’s; biomarkers; single-molecule.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Aged, 80 and over
  • Amyloidogenic Proteins / cerebrospinal fluid*
  • Benzothiazoles
  • Circular Dichroism / methods
  • Diagnostic Imaging / methods*
  • Female
  • Humans
  • Male
  • Microscopy, Atomic Force / methods
  • Microscopy, Fluorescence / methods*
  • Middle Aged
  • Parkinson Disease / cerebrospinal fluid*
  • Thiazoles

Substances

  • Amyloidogenic Proteins
  • Benzothiazoles
  • Thiazoles
  • thioflavin T