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Article

Aequorin as a Useful Calcium-Sensing Reporter in Candida albicans

Institute of Microbiology, University of Lausanne and University Hospital, CH-1011 Lausanne, Switzerland
Academic Editor: Juliana Campos Junqueira
J. Fungi 2021, 7(4), 319; https://doi.org/10.3390/jof7040319
Received: 29 January 2021 / Revised: 14 April 2021 / Accepted: 19 April 2021 / Published: 20 April 2021
(This article belongs to the Special Issue New Perspectives for Candidiasis)
In Candida albicans, calcium ions (Ca2+) regulate the activity of several signaling pathways, especially the calcineurin signaling pathway. Ca2+ homeostasis is also important for cell polarization, hyphal extension, and plays a role in contact sensing. It is therefore important to obtain accurate tools with which Ca2+ homeostasis can be addressed in this fungal pathogen. Aequorin from Aequorea victoria has been used in eukaryotic cells for detecting intracellular Ca2+. A codon-adapted aequorin Ca2+-sensing expression system was therefore designed for probing cytosolic Ca2+ flux in C. albicans. The availability of a novel water-soluble formulation of coelenterazine, which is required as a co-factor, made it possible to measure bioluminescence as a readout of intracellular Ca2+ levels in C. albicans. Alkaline stress resulted in an immediate influx of Ca2+ from the extracellular medium. This increase was exacerbated in a mutant lacking the vacuolar Ca2+ transporter VCX1, thus confirming its role in Ca2+ homeostasis. Using mutants in components of a principal Ca2+ channel (MID1, CCH1), the alkaline-dependent Ca2+ spike was greatly reduced, thus highlighting the crucial role of this channel complex in Ca2+ uptake and homeostasis. Exposure to the antiarrhythmic drug amiodarone, known to perturb Ca2+ trafficking, resulted in increased cytoplasmic Ca2+ within seconds that was abrogated by the chelation of Ca2+ in the external medium. Ca2+ import was also dependent on the Cch1/Mid1 Ca2+ channel in amiodarone-exposed cells. In conclusion, the aequorin Ca2+ sensing reporter developed here is an adequate tool with which Ca2+ homeostasis can be investigated in C. albicans. View Full-Text
Keywords: Candida; aequorin; calcium homeostasis; alkaline stress; amiodarone Candida; aequorin; calcium homeostasis; alkaline stress; amiodarone
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MDPI and ACS Style

Sanglard, D. Aequorin as a Useful Calcium-Sensing Reporter in Candida albicans. J. Fungi 2021, 7, 319. https://doi.org/10.3390/jof7040319

AMA Style

Sanglard D. Aequorin as a Useful Calcium-Sensing Reporter in Candida albicans. Journal of Fungi. 2021; 7(4):319. https://doi.org/10.3390/jof7040319

Chicago/Turabian Style

Sanglard, Dominique. 2021. "Aequorin as a Useful Calcium-Sensing Reporter in Candida albicans" Journal of Fungi 7, no. 4: 319. https://doi.org/10.3390/jof7040319

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