FRET and flow cytometry assays to measure proteopathic seeding activity in biological samples

Jennifer L. Furman, Marc I. Diamond

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

Transcellular propagation of protein aggregates—or seeds—is increasingly implicated as a mechanism for disease progression in many neurodegenerative disorders, including Alzheimer’s disease and the related tauopathies. While neuropathology generally originates in one discrete brain region, pathology progresses as disease severity advances, often along discrete neural networks. The stereotypical spread of tau pathology suggests that cell-to-cell transfer of toxic protein aggregates could underlie disease progression, and recent studies implicate seeding as a proximal marker of disease, as compared to standard histological and biochemical analyses. Commonly used metrics for protein aggregation detection, however, lack sensitivity, are not quantitative, and/or undergo subjective classification. Here, we describe a FRET and flow cytometry cell-based assay that allows for rapid and quantitative detection of protein aggregates from human and rodent biological specimens.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages349-359
Number of pages11
Volume1523
DOIs
StatePublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1523
ISSN (Print)10643745

Keywords

  • Flow cytometry
  • FRET
  • Huntingtin
  • Mouse model
  • Neurodegeneration
  • Prion
  • Protein aggregation
  • Spread
  • Synuclein
  • Tau

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Furman, J. L., & Diamond, M. I. (2017). FRET and flow cytometry assays to measure proteopathic seeding activity in biological samples. In Methods in Molecular Biology (Vol. 1523, pp. 349-359). (Methods in Molecular Biology; Vol. 1523). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6598-4_23