Journal of Bioenergetics and Biomembranes

, Volume 49, Issue 3, pp 281–290 | Cite as

Acridine yellow. A novel use to estimate and measure the plasma membrane potential in Saccharomyces cerevisiae

  • Martha Calahorra
  • Norma Silvia Sánchez
  • Antonio Peña
Article

Abstract

Translocation of ions and other molecules across the plasma membrane of yeast requires the electric potential generated by a H+-ATPase. We measured under different conditions fluorescence changes and accumulation of acridine yellow, looking for qualitative and quantitative estimations of the PMP in Saccharomyces cerevisiae in various conditions. Fluorescence changes indicated an accumulation of the dye requiring a substrate, and accumulation and quenching by mitochondria that could be released by an uncoupler. K+ produced a decrease of the fluorescence that was much lower upon the addition of Na+. These changes were confirmed by images of the cells under the microscope. The dye accumulation under different conditions showed changes consistent with the physiological situation of the cells. Since it accumulates due to the PMP, but a large part of it binds to the internal components, we permeabilized the cells with chitosan to subtract this factor and correct the accumulation data. Both raw and corrected values of PMP are different to those obtained before by other authors and our group, showing acridine yellow as a promising indicator to follow changes of the PMP by the fluorescence changes, but also by its accumulation. Under conditions described, the dye is a low cost monitor to define and follow qualitative and quantitative changes of PMP in yeast. Acridine yellow can also be used to follow changes of the mitochondrial membrane potential.

Keywords

Organic cations Plasma membrane potential Yeast Acridine yellow 

Abbreviations

AcYe

Acridine yellow

CCCP

Carbonylcyanide-m- phenylhydrazine

DiSC3(3)

3,3-dipropylthiacarbocyanine

MES

Morpholinoethanesulphonic acid

PMP

Plasma membrane electric potential difference

TEA

Triethanolamine.

Notes

Acknowledgements

The authors thank Dr.Yazmín Ramiro Cortés of our Institute for her invaluable help with the microscope images. Authors thank Juan Manuel Barbosa and Ivett Rosas from our computing unit, for their invaluable assistance. This work was supported by grants 238497 from the Consejo Nacional de Ciencia y Tecnología, México, and grants IN223999, and IN202103 from the Universidad Nacional Autónoma de México.

Compliance with ethical standards

Competing interests

The authors have declared that no conflict of interest exist.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Departamento de Genética Molecular, Instituto de Fisiología CelularUniversidad Nacional Autónoma de MéxicoMexico CityMexico

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