Exploiting flow cytometry for the unbiased quantification of protein inclusions in Caenorhabditis elegans

Kristian Claesson, Yee Lian Chew, Heath Ecroyd

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Abstract

The aggregation of proteins into inclusions or plaques is a prominent hallmark of a diverse range of pathologies including neurodegenerative diseases. The quantification of such inclusions in Caenorhabditis elegans models of aggregation is usually achieved by fluorescence microscopy or other techniques involving biochemical fractionation of worm lysates. Here, we describe a simple and rapid flow cytometry-based approach that allows fluorescently tagged inclusions to be enumerated in whole worm lysate in a quantitative and unbiased fashion. We demonstrate that this technique is applicable to multiple C. elegans models of aggregation and importantly, can be used to monitor the dynamics of inclusion formation in response to heat shock and during ageing. This includes the characterisation of physicochemical properties of inclusions, such as their apparent size, which may reveal how aggregate formation is distinct in different tissues or at different stages of pathology or ageing. This new method can be used as a powerful technique for the medium- to high-throughput quantification of inclusions in future studies of genetic or chemical modulators of aggregation in C. elegans. (Figure presented.)

Original languageEnglish
Pages (from-to)281-292
Number of pages12
JournalJournal of Neurochemistry
Volume161
Issue number3
Early online date15 Feb 2022
DOIs
Publication statusPublished - May 2022

Keywords

  • amyotrophic lateral sclerosis
  • C. elegans
  • flow cytometry
  • Huntington's disease
  • inclusions
  • protein aggregation

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