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The Use of Thioflavin T for the Estimation and Measurement of the Plasma Membrane Electric Potential Difference in Different Yeast Strains

The use of the cationic, dye thioflavin T (ThT), to estimate the electric plasma membrane potential difference (PMP) via the fluorescence changes and to obtain its actual values from the accumulation of the dye, considering important correction factors by its binding to the internal components of th...

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Autores principales: Peña, Antonio, Sánchez, Norma Silvia, Padilla-Garfias, Francisco, Ramiro-Cortés, Yazmín, Araiza-Villanueva, Minerva, Calahorra, Martha
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10532974/
https://www.ncbi.nlm.nih.gov/pubmed/37755056
http://dx.doi.org/10.3390/jof9090948
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author Peña, Antonio
Sánchez, Norma Silvia
Padilla-Garfias, Francisco
Ramiro-Cortés, Yazmín
Araiza-Villanueva, Minerva
Calahorra, Martha
author_facet Peña, Antonio
Sánchez, Norma Silvia
Padilla-Garfias, Francisco
Ramiro-Cortés, Yazmín
Araiza-Villanueva, Minerva
Calahorra, Martha
author_sort Peña, Antonio
collection PubMed
description The use of the cationic, dye thioflavin T (ThT), to estimate the electric plasma membrane potential difference (PMP) via the fluorescence changes and to obtain its actual values from the accumulation of the dye, considering important correction factors by its binding to the internal components of the cell, was described previously for baker’s yeast. However, it was considered important to explore whether the method developed could be applied to other yeast strains. Alternative ways to estimate the PMP by using flow cytometry and a multi-well plate reader are also presented here. The methods were tested with other strains of Saccharomyces cerevisiae (W303-1A and FY833), as well as with non-conventional yeasts: Debaryomyces hansenii, Candida albicans, Meyerozyma guilliermondii, and Rhodotorula mucilaginosa. Results of the estimation of the PMP via the fluorescence changes under different conditions were adequate with all strains. Consistent results were also obtained with several mutants of the main monovalent transporters, validating ThT as a monitor for PMP estimation.
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spelling pubmed-105329742023-09-28 The Use of Thioflavin T for the Estimation and Measurement of the Plasma Membrane Electric Potential Difference in Different Yeast Strains Peña, Antonio Sánchez, Norma Silvia Padilla-Garfias, Francisco Ramiro-Cortés, Yazmín Araiza-Villanueva, Minerva Calahorra, Martha J Fungi (Basel) Article The use of the cationic, dye thioflavin T (ThT), to estimate the electric plasma membrane potential difference (PMP) via the fluorescence changes and to obtain its actual values from the accumulation of the dye, considering important correction factors by its binding to the internal components of the cell, was described previously for baker’s yeast. However, it was considered important to explore whether the method developed could be applied to other yeast strains. Alternative ways to estimate the PMP by using flow cytometry and a multi-well plate reader are also presented here. The methods were tested with other strains of Saccharomyces cerevisiae (W303-1A and FY833), as well as with non-conventional yeasts: Debaryomyces hansenii, Candida albicans, Meyerozyma guilliermondii, and Rhodotorula mucilaginosa. Results of the estimation of the PMP via the fluorescence changes under different conditions were adequate with all strains. Consistent results were also obtained with several mutants of the main monovalent transporters, validating ThT as a monitor for PMP estimation. MDPI 2023-09-20 /pmc/articles/PMC10532974/ /pubmed/37755056 http://dx.doi.org/10.3390/jof9090948 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Peña, Antonio
Sánchez, Norma Silvia
Padilla-Garfias, Francisco
Ramiro-Cortés, Yazmín
Araiza-Villanueva, Minerva
Calahorra, Martha
The Use of Thioflavin T for the Estimation and Measurement of the Plasma Membrane Electric Potential Difference in Different Yeast Strains
title The Use of Thioflavin T for the Estimation and Measurement of the Plasma Membrane Electric Potential Difference in Different Yeast Strains
title_full The Use of Thioflavin T for the Estimation and Measurement of the Plasma Membrane Electric Potential Difference in Different Yeast Strains
title_fullStr The Use of Thioflavin T for the Estimation and Measurement of the Plasma Membrane Electric Potential Difference in Different Yeast Strains
title_full_unstemmed The Use of Thioflavin T for the Estimation and Measurement of the Plasma Membrane Electric Potential Difference in Different Yeast Strains
title_short The Use of Thioflavin T for the Estimation and Measurement of the Plasma Membrane Electric Potential Difference in Different Yeast Strains
title_sort use of thioflavin t for the estimation and measurement of the plasma membrane electric potential difference in different yeast strains
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10532974/
https://www.ncbi.nlm.nih.gov/pubmed/37755056
http://dx.doi.org/10.3390/jof9090948
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