Imaging polyphenolic therapeutic compounds in a eukaryotic model microbe
Ferrara, Billy and Thompson, Elinor ORCID: https://orcid.org/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) |
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Additional Information: | Microbiology Society Annual Conference 2018 held from 10–13 April, at the ICC Birmingham, UK. |
Uncontrolled Keywords: | flavonoid, Dictyostelium, fluorescence imaging |
Faculty / School / Research Centre / Research Group: | Faculty of Engineering & Science Faculty of Engineering & Science > Biology & Biotechnology Research Group Faculty of Engineering & Science > School of Science (SCI) Faculty of Education, Health & Human Sciences > School of Human Sciences (HUM) |
Last Modified: | 09 Oct 2021 04:45 |
URI: | http://gala.gre.ac.uk/id/eprint/19661 |
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