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

Journal Article


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.).

UOW Authors


Publication Date


  • 2022

Citation


  • Claesson, K., Chew, Y. L., & Ecroyd, H. (2022). Exploiting flow cytometry for the unbiased quantification of protein inclusions in Caenorhabditis elegans. Journal of Neurochemistry, 161(3), 281-292. doi:10.1111/jnc.15591

Scopus Eid


  • 2-s2.0-85125547703

Start Page


  • 281

End Page


  • 292

Volume


  • 161

Issue


  • 3

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.).

UOW Authors


Publication Date


  • 2022

Citation


  • Claesson, K., Chew, Y. L., & Ecroyd, H. (2022). Exploiting flow cytometry for the unbiased quantification of protein inclusions in Caenorhabditis elegans. Journal of Neurochemistry, 161(3), 281-292. doi:10.1111/jnc.15591

Scopus Eid


  • 2-s2.0-85125547703

Start Page


  • 281

End Page


  • 292

Volume


  • 161

Issue


  • 3