Shining a light on the impact of antifungals on Aspergillus fumigatus subcellular dynamics through fluorescence imaging

Storer, I. S. R., Sastré-Velásquez, L. E., Easter, T., Mertens, B., Dallemulle, A., Bottery, M., Tank, R., Offterdinger, M., Bromley, M. J., van Rhijn, N. and Gsaller, F. (2024) Shining a light on the impact of antifungals on Aspergillus fumigatus subcellular dynamics through fluorescence imaging. Antimicrobial Agents and Chemotherapy, 68 (11). ISSN 0066-4804

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Abstract

Fluorescent proteins (FPs) are indispensable tools used for molecular imaging, single-cell dynamics, imaging in infection models, and more. However, next-generation FPs have yet to be characterized in Aspergillus. Here, we characterize 18 FPs in the pathogenic filamentous fungus Aspergillus fumigatus spanning the visible light spectrum. We report on in vivo FP brightness in hyphal and spore morphotypes and show how a fluoropyrimidine-based selection system can be used to iteratively introduce four distinct FPs enabling the simultaneous visualization of the cell membrane, mitochondria, peroxisomes, and vacuoles. Using this strain, we describe and compare the dynamic responses of organelles to stresses induced by voriconazole, amphotericin B, and the novel antifungal drugs olorofim and manogepix. The expansion to the fluorescent genetic toolbox will overcome boundaries in research applications that involve fluorescence imaging in filamentous fungi.

Item Type:	Article
Additional Information:	Funding information: This research was funded in whole or in part by the Austrian Science Fund (FWF) (grant DOI: 10.55776/P35951 and DOI: 10.55776/P31093) to F.G. L.E.S.-V. participated in the HOROS program W1253 funded by the FWF. I.S.R.S. was supported by MRC grant reference MR/N013751/1. N.V.R. was supported by the Wellcome Trust (grant number: 226408/Z/22/Z). M.J.B. was supported by Wellcome trust project number 219551/Z/19/Z. Rights Retention Statement: For open access purposes, the author has applied a CC BY public copyright license to any author-accepted manuscript version arising from this submission.
Uncontrolled Keywords:	antifungal agents,aspergillus,aspergillus fumigatus,fluorescence,fluorophores,fungal disease,imaging,microscopy,pharmacology,pharmacology (medical),infectious diseases,sdg 3 - good health and well-being ,/dk/atira/pure/subjectarea/asjc/3000/3004
Faculty \ School:	Faculty of Science > School of Chemistry, Pharmacy and Pharmacology
Related URLs:	http://www.scopus.com/inward/record.url?...
Depositing User:	LivePure Connector
Date Deposited:	20 Aug 2025 14:30
Last Modified:	27 Aug 2025 08:28
URI:	https://ueaeprints.uea.ac.uk/id/eprint/100176
DOI:	10.1128/aac.00803-24

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