Abstract
Digital image analysis of high time resolution video microscopy was used to investigate hyphal growth dynamics in different Candida albicans strains. The effects of the quorum sensing molecules tyrosol and farnesol, the deletion of the fungus specific protein phosphatase Z1 CaPPZ1), and the hypha-specific cyclin (HGC1) genes were analyzed by this method. Our system monitored cell growth in a CO2 incubator under near-physiological conditions and measured three major parameters under the following stringent conditions: (a) the time of yeast cell adherence, (b) the time of hyphal outgrowth, and (c) the rate of hyphal growth. This method showed that hyphal extension of wild-type SC5314 cells was accelerated by tyrosol and inhibited by farnesol. Hyphal growth rate was moderately lower in cappz1 and strongly reduced in hgc1 mutants. In addition, tyrosol treatment caused a firm adherence, while farnesol treatment and hgc1 mutation prevented the adherence of yeast cells to the surface of the culture flask. Transition from yeast-to-hyphal state was faster after tyrosol treatment, while it was reduced in farnesol-treated cells as well as in the cappz1 and hgc1 mutants. Our data confirm the notion that the attachment of yeast cells, the yeast-to-hyphal transition, and hyphal growth rate are closely related processes. Time-lapse video microscopy combined with image analysis offers a convenient and reliable method of testing chemicals, including potential drug candidates, and genetic manipulations on the dynamic morphological changes in C. albicans strains.
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Acknowledgements
Thanks are due to Prof. Alexander Johnson (Department of Biochemistry and Biophysics, University of California, San Francisco, USA) for the QMY23 and to Prof. Yue Wang (Institute of Molecular and Cell Biology, Singapore, Singapore) for the WYZ12.1 and WYZ12.2C. albicans strains. This work was supported by the OTKA grant K108989 and by the UD Faculty of Medicine Research Fund (Bridging Fund 2012) to VD, TÁMOP 4.2.1/B-09/1/KONV-2010-0007 to IP, as well as by TÁMOP 4.2.4. A/2-11-1-2012-0001 National Excellence Program that was subsidized by the European Union and co-financed by the European Social Fund to GN and KP. GWH is supported by grants from the National Center for Research Resources (5P20RR018751-09) and the National Institute of General Medical Sciences (8 P20 GM103513-09) from the National Institutes of Health.
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Nagy, G., Hennig, G.W., Petrenyi, K. et al. Time-lapse video microscopy and image analysis of adherence and growth patterns of Candida albicans strains. Appl Microbiol Biotechnol 98, 5185–5194 (2014). https://doi.org/10.1007/s00253-014-5696-5
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DOI: https://doi.org/10.1007/s00253-014-5696-5
Keywords
- Candida albicans
- Quorum-sensing molecules
- Genetic mutations
- Yeast cell adherence
- Hyphae outgrowth
- Image analysis