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Imaging polyphenolic therapeutic compounds in a eukaryotic model microbe

Imaging polyphenolic therapeutic compounds in a eukaryotic model microbe

Ferrara, Billy and Thompson, Elinor ORCID: 0000-0002-6434-9290 (2018) Imaging polyphenolic therapeutic compounds in a eukaryotic model microbe. In: Microbiology Society Annual Conference 2018.

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Abstract

Flavonoids are polyphenolic metabolites that have a range of physiological and developmental functions in plants. They are the focus of much work as potential therapeutics, although investigation of specific mode of action remains a notably under-researched area. Monitoring transport and location of flavonoids in cells is difficult because, despite a role in UV-absorption in plants, they emit only low levels of fluorescence. Visualising them in plants is possible using the Naturstoff reagent (NA), reported historically to be a polyphenol-fluorescence-enhancing stain. We explored therefore whether this agent was effective during preclinical assessment of polyphenolic therapeutics in a microbial-model.

The eukaryote Dictyostelium discoideum has been shown to be a useful model when identifying novel drug targets for treating various diseases. For example, in the case of polycystic kidney disease, naringenin decreased Dictyostelium cell division whereas a polycystin-2-null Dictyostelium line was resistant to the flavonoid, and, subsequently, naringenin treatment proved to reduce cyst-formation in mammalian-kidney model cell lines1. To monitor transport and site of action of the drugs investigated in such studies, we developed a method using NA-staining in this model organism. A range of polyphenolics were assayed in cells, cell-extracts and the cell-washes, and NA-enhanced imaging was evaluated in parallel with LCMS-quantification. NA-enhanced fluorescence of compounds at therapeutically relevant concentrations proved an effective and qualitative measure of transport and localisation in Dictyostelium, and could be used in concert with localisation dyes. Fluorescence-enhancement is limited to a subset of flavonoids, however, and not more widely applicable in our studies to date.

Item Type: Conference or Conference Paper (Poster)
Additional Information: Microbiology Society Annual Conference 2018 held from 10–13 April, at the ICC Birmingham, UK.
Uncontrolled Keywords: flavonoid, Dictyostelium, fluorescence imaging
Faculty / Department / Research Group: Faculty of Engineering & Science
Faculty of Engineering & Science > Biology & Biotechnology Research Group
Faculty of Engineering & Science > Department of Life & Sports Sciences
Faculty of Engineering & Science > Department of Pharmaceutical, Chemical & Environmental Sciences
Last Modified: 07 Jun 2019 15:43
Selected for GREAT 2016: None
Selected for GREAT 2017: None
Selected for GREAT 2018: None
Selected for GREAT 2019: None
URI: http://gala.gre.ac.uk/id/eprint/19661

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