Microtubules and Actin Cytoskeleton of Potentially Pathogenic Basidiomycetous Yeast as Targets for Antifungals

Kopecká, Marie; Gabriel, Miroslav

doi:10.1159/000224655

Cited by 15 publications

(20 citation statements)

References 51 publications

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“…To obtain an exponential culture, cells were cultivated in YEPD medium [1% (w/v) yeast extract, 1% (w/v) peptone and 1% (w/v) glucose] on a shaker overnight at 23°C (16 h) and were then diluted to 5 × 10 5 to 1 × 10 6 cells/ml by 1% YEPD medium and pipetted in 500-μl volumes to test tubes. They were then used for the application of inhibitors and shaken in the dark in a water bath for 6 or 14 days [12,14]. …”

Section: Methodsmentioning

confidence: 99%

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Effects of Microtubule and Actin Inhibitors on Cryptococcus neoformans Examined by Scanning and Transmission Electron Microscopy

Kopecká

2014

Chemotherapy

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Background:Cryptococcus neoformans is one of the most important human fungal pathogens. Its cells contain rich microtubules required for nuclear division and rich F-actin cytoskeletons for cell division. Disruption of microtubules by a microtubule inhibitor should block nuclear division, and disruption of F-actin by an actin inhibitor should block cell division. We investigated the effects of microtubule and actin inhibitors to find out whether the cytoskeletons of C. neoformans can become a new anti-fungal target for the inhibition of cell division, when examined at the ultrastructural level. Methods: Cells treated with the microtubule inhibitors vincristine (VIN) and methyl benzimidazole-2-ylcarbamate (BCM) and the actin inhibitor latrunculin A (LA), in yeast extract peptone dextrose medium, were examined by scanning (SEM) and transmission electron microscopy (TEM), and the cell number was counted using a Bürker chamber. Results: After 2 days of inhibition with VIN, BCM or LA, the cells did not divide, but later, resistant, proliferating cells appeared in all samples. With combined microtubule and actin inhibitors (VIN + LA or BCM + LA), cells did not divide during 6 or even 14 days, and no resistant cells originated. TEM showed that the inhibited cells were without cytoplasm and were dead; only empty cell walls persisted with reduced capsules, shown on SEM. Conclusion: Combined microtubule and actin inhibitors (VIN + LA or BCM + LA), have lethal effects on C. neoformans cells and no resistant cells originate.

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Section: Methodsmentioning

confidence: 99%

“…10 m M of stock solution was prepared by dissolving 100 μg of LA in 25 μl of dimethyl sulfoxide (DMSO), kept at -20°C [12,14]. …”

Section: Methodsmentioning

confidence: 99%

Effects of Microtubule and Actin Inhibitors on Cryptococcus neoformans Examined by Scanning and Transmission Electron Microscopy

Kopecká

2014

Chemotherapy

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“…10 m M stock solution was prepared by dissolving 100 μg of LA (Molecular Probes, USA) in 25 μl of DMSO, kept at −20°C and then added to cells in 1% YEPD to a final concentration of 100 μ M that contained 1% DMSO [7,8,10,11]; 500-μl volumes of cultures in test tubes were shaken in the dark in a water bath. Samples for EM were taken before the LA treatment, i.e.…”

Section: Methodsmentioning

confidence: 99%

The Effects of the F-Actin Inhibitor Latrunculin A on the Pathogenic Yeast Cryptococcus neoformans

2014

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Background: This basic research aimed to investigate the effects of the actin inhibitor latrunculin A (LA) on the human pathogen Cryptococcus neoformans, by freeze-substitution (FS) and electron microscopy (EM), to determine whether the actin cytoskeleton can become a new antifungal target for inhibition of cell division. Methods: Cells treated with LA for 20 h in yeast-extract peptone dextrose medium were investigated by phase-contrast and fluorescent microscopy, FS and transmission EM, counted in a Bürker chamber and the absorbance was then measured. Results: The disappearance of actin patches, actin cables and actin rings demonstrated the response of the cells of C. neoformans to the presence of the actin inhibitor LA. The removal of actin cables and patches arrested proliferation and led to the production of cells that had ultrastructural disorder, irregular morphology of the mitochondria and thick aberrant cell walls. Budding cells lysed in the buds and septa. Conclusion: LA exerts fungistatic, fungicidal and fungilytic effects on the human pathogenic yeast C. neoformans.

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“…Moreover, the problem of drug resistance through overexpression of efflux pumps in fungal infections and in cancer chemotherapy is similar. Many of the targets for antineoplastic drugs have homologs in yeast and the eukaryotic fungal pathogens including C. albicans [17,18]. Huge efforts are being made worldwide to synthesize molecules for the prevention and treatment of cancer, which have resulted in a large variety of candidate molecules.…”

Section: Introductionmentioning

confidence: 99%

Dual Properties of Anticancer Agents: An Exploratory Study on the in vitro Anti-Candida Properties of Thirty Drugs

Routh

Raut

Karuppayil³

2011

Chemotherapy

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Background: Anticancer/antineoplastic agents could be a good resource for the discovery of novel antifungal agents and targets since human beings share a common eukaryotic heritage with fungi. Methods: Thirty commonly prescribed anticancer drugs belonging to 12 different classes were analyzed for their effects on the growth of Candida albicans. Minimum inhibitory concentrations (MICs) were obtained using standard CLSI-M27 A2 methodology, and minimal fungicidal concentrations (MFCs) were determined via the agar plate method. Results: Anticancer agents inhibited the growth of C. albicans in a concentration-dependent manner. Nine drugs from different classes were effective at low concentrations (≤50 µg·ml^–1), while 15 anticancer drugs exhibited MICs of 100 µg·ml^–1. Sixteen out of 30 drugs exerted fungicidal activity in the range of 400–800 µg·ml^–1. Conclusions: MICs and MFCs for 30 anticancer drugs were established against C. albicans. Our study highlighted the anti-Candida potential of these drugs, which may give insights to unexplored targets for antifungal chemotherapy.

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Microtubules and Actin Cytoskeleton of Potentially Pathogenic Basidiomycetous Yeast as Targets for Antifungals

Cited by 15 publications

References 51 publications

Effects of Microtubule and Actin Inhibitors on <b><i>Cryptococcus neoformans</i></b> Examined by Scanning and Transmission Electron Microscopy

Effects of Microtubule and Actin Inhibitors on <b><i>Cryptococcus neoformans</i></b> Examined by Scanning and Transmission Electron Microscopy

The Effects of the F-Actin Inhibitor Latrunculin A on the Pathogenic Yeast <b><i>Cryptococcus neoformans</i></b>

Dual Properties of Anticancer Agents: An Exploratory Study on the in vitro Anti-Candida Properties of Thirty Drugs

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