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An Ex vivo Assay to Study Candida albicans Hyphal Morphogenesis in the Gastrointestinal Tract
Published on: July 1, 2020
Changes in the Ultrastructure of Candida albicans Treated with Cationic Peptides
Alina Grigor'eva1, Alevtina Bardasheva1, Anastasiya Tupitsyna1
1Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Science, Lavrent'ev av., 8, 630090 Novosibirsk, Russia.
Abstract:
Candida albicans is becoming increasingly harmful for humans, which determines the need for new effective antifungal preparations. Currently, when testing antifungals, various morphological methods are used, among which transmission electron microscopy (TEM) is not the leading one. In this work, we used TEM to study the submicroscopic changes in C. albicans cells induced by cationic peptides R9F2 and (KFF)3K. Studies were performed on C. albicans-34 strain from the Collection of EMTC of ICBFM SB RAS in logarithmic phase. R9F2 and (KFF)3K showed an antifungal effect (MIC 10 and 20 μM) and suppressed fungal hyphal growth. Semithin and ultrathin sections of fungal suspensions incubated with 10 μM of peptides were studied at regular intervals from 15 min to 24 h. The first target of both peptides was plasmalemma, and its "alignment" was the only common morphological manifestation of their effect. Other changes in the plasmalemma and alteration of the vacuole and cell wall ultrastructure distinctly differed in cells treated with R9F2 and (KFF)3K peptides. In general, our work has shown pronounced differences of the temporal and morphologic characteristics of the effect of peptides, evidently related to their physicochemical properties. The benefit of TEM studies of ultrathin sections for understanding the mechanisms of action of antifungal drugs is shown.
Insights
New cationic peptides R9F2 and (KFF)3K show antifungal effects against Candida albicans. Transmission electron microscopy revealed distinct submicroscopic changes in fungal cells, highlighting TEM
Area of Science:
- Mycology
- Biochemistry
- Microbiology
Background:
- Candida albicans poses a growing threat, necessitating novel antifungal agents.
- Morphological methods are used to test antifungals, but transmission electron microscopy (TEM) is underutilized.
- Cationic peptides are a promising class of antimicrobial compounds.
Purpose of the Study:
- To investigate the submicroscopic effects of cationic peptides R9F2 and (KFF)3K on Candida albicans using TEM.
- To compare the morphological changes induced by R9F2 and (KFF)3K.
- To demonstrate the utility of TEM in understanding antifungal drug mechanisms.
Main Methods:
- Candida albicans-34 strain was cultured to the logarithmic phase.
- Cells were treated with cationic peptides R9F2 and (KFF)3K at sub-inhibitory concentrations (10 μM).
- Submicroscopic changes were analyzed using transmission electron microscopy (TEM) on semithin and ultrathin sections at various time points (15 min to 24 h).
Main Results:
- Both R9F2 and (KFF)3K exhibited antifungal activity, inhibiting growth at 10 and 20 μM respectively, and suppressing hyphal formation.
- The primary target for both peptides was the plasmalemma, with "alignment" being the sole common ultrastructural change observed.
- Distinct differences were noted in other plasmalemma alterations and in the ultrastructure of the vacuole and cell wall between R9F2 and (KFF)3K treated cells.
- Temporal and morphological characteristics of peptide effects varied significantly, likely due to physicochemical differences.
Conclusions:
- TEM provides valuable insights into the mechanisms of action of antifungal peptides.
- R9F2 and (KFF)3K exert differential effects on Candida albicans ultrastructure.
- The study underscores the potential of cationic peptides as antifungal agents and TEM as a key analytical tool.
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