Abstract
The potential of the plasma membrane (Δψ) regulates the electrochemical potential between the outer and inner sides of cell membranes. The opportunistic fungal pathogen, Candida albicans, regulates the membrane potential in response to environmental conditions, as well as the physiological state of the cell. Here we demonstrate a new method for detection of cell membrane depolarization/permeabilization in C. albicans using the potentiometric zwitterionic dye di-4-ANEPPS. Di-4-ANEPPS measures the changes in the cell Δψ depending on the phases of growth and external factors regulating Δψ, such as potassium or calcium chlorides, amiodarone or DM-11 (inhibitor of H+-ATPase). We also demonstrated that di-4-ANEPPS is a good tool for fast measurement of the influence of amphipathic compounds on Δψ.
Highlights
Though the plasma membrane potential (∆ψ) is an electrochemical potential difference between extracellular and intracellular compartments in all living cells, the mechanisms maintaining ∆ψ differ between cell types [1]. ∆ψ acts as an indicator of the physiological status of the cell; for example, depolarization of the cell membrane in lymphocytes prevents cell proliferation [2]
Our previous investigations indicated that the ∆ψ of C. albicans measured by diS-C3(3) dye differs from S. cerevisiae [15]
We reported the use of di-4-ANEPPS dye on Δѱ measurements of C. albicans
Summary
Though the plasma membrane potential (∆ψ) is an electrochemical potential difference between extracellular and intracellular compartments in all living cells, the mechanisms maintaining ∆ψ differ between cell types [1]. ∆ψ acts as an indicator of the physiological status of the cell; for example, depolarization of the cell membrane in lymphocytes prevents cell proliferation [2]. The influence of the value of ∆ψ on the lipid lateral localization in the plasma membrane of the yeast Saccharomyces cerevisiae is another example that highlights the importance of the ∆ψ in cell biology [3]. The value of C. albicans ∆ψ is ~−120 mV and is comparable to that of pathogenic bacteria, which ranges from ~−130 mV to ~−150 mV [5,6]. Unlike in C. albicans, the value of ∆ψ of non-pathogenic S. cerevisiae is ~−71 mV and is comparable to the potential of mammalian cells, which is ~−90 mV [5,6]. The loss of cell membrane integrity due to the action of antifungal drugs causes plasma membrane depolarization [9]
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