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Analysis of the Development of a Morphological Phenotype as a Function of Protein Concentration in Budding Yeast
Published on: March 24, 2010
The link between yeast cell wall porosity and plasma membrane permeability after PEF treatment
Arunas Stirke1, Raimonda Celiesiute-Germaniene2, Aurelijus Zimkus3
1Center for Physical Sciences and Technology, Sauletekio ave. 3, LT-10257, Vilnius, Lithuania.
Abstract:
An investigation of the yeast cell resealing process was performed by studying the absorption of the tetraphenylphosphonium (TPP+) ion by the yeast Saccharomyces cerevisiae. It was shown that the main barrier for the uptake of such TPP+ ions is the cell wall. An increased rate of TPP+ absorption after treatment of such cells with a pulsed electric field (PEF) was observed only in intact cells, but not in spheroplasts. The investigation of the uptake of TPP+ in PEF treated cells exposed to TPP+ for different time intervals also showed the dependence of the absorption rate on the PEF strength. The modelling of the TPP+ uptake recovery has also shown that the characteristic decay time of the non-equilibrium (PEF induced) pores was approximately a few tens of seconds and this did not depend on the PEF strength. A further investigation of such cell membrane recovery process using a florescent SYTOX Green nucleic acid stain dye also showed that such membrane resealing takes place over a time that is like that occurring in the cell wall. It was thus concluded that the similar characteristic lifetimes of the non-equilibrium pores in the cell wall and membrane after exposure to PEF indicate a strong coupling between these parts of the cell.
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