Journal of Bioenergetics and Biomembranes

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

RETRACTED ARTICLE: Acridine yellow.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

  • Martha CalahorraEmail author
  • Norma Silvia Sánchez
  • Antonio Peña


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.


Organic cations Plasma membrane potential Yeast Acridine yellow 



Acridine yellow


Carbonylcyanide-m- phenylhydrazine




Morpholinoethanesulphonic acid


Plasma membrane electric potential difference





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.


  1. Azzi A, Chance B, Radda GK, Lee CP (1969) A fluorescence probe of energy-dependent structure changes in fragmented membranes. Proc Natl Acad Sci USA Feb 62(2):612–619CrossRefGoogle Scholar
  2. Boxman AW, Barts PWJA, Borst-Pauwels GWFH (1982) Some characteristics of tetraphenylphosphonium uptake into Saccharomyces cerevisiae. Biochim Biophys Acta 686:13–18CrossRefGoogle Scholar
  3. Gásková D, Brodska B, Herman P, Vecer J, Malinsky J, Sigler K, Benada O, Plasek O (1998) Fluorescent probing of membrane potential in walled cells: diS-C3(3) assay in Saccharomyces cerevisiae. Yeast 14:1189–1197CrossRefPubMedCentralGoogle Scholar
  4. Gitler C, Rubalcava B, Caswell A (1969) Fluorescence changes of ethidium bromide on binding to erythrocyte and mitochondrial membranes. Biochim Biophys Acta 193(2):479–481CrossRefGoogle Scholar
  5. Kinclova-Zimmermannova O, Gásková D, Sychrova H (2006) The Na+, K+/H+ antiporter influences the plasma membrane potential of Saccharomyces cerevisiae. FEMS Yeast Res 6:792–800CrossRefPubMedCentralGoogle Scholar
  6. Kotyk A (1963) Intracellular pH of Baker's yeast. Folia Microbiol 8:27–31CrossRefGoogle Scholar
  7. López R, Enríquez E, Peña A (1999) Effects of weak acids on cation accumulation, ΔpH and ΔΨ in yeast. Yeast 15:553–562CrossRefPubMedCentralGoogle Scholar
  8. Maresova L, Urbankova E, Gásková D, Sychrova H (2006) Measurements of plasma membrane potential changes in Saccharomyces cerevisiae cells reveal the importance of the Tok1 channel in membrane potential maintenance. FEMS Yeast Res 6(7):1039–1046CrossRefPubMedCentralGoogle Scholar
  9. Maresova L, Muend S, Zhang Y-Q, Sychrova H, Rao R (2009) Membrane hyperpolarization drives cation influx and fungicidal activity of amiodarone. J Biol Chem 284:2795–2802CrossRefPubMedCentralGoogle Scholar
  10. Peña A, Ramírez G (1975) Interaction of ethidium bromide with the transport system for monovalent cations in yeast. J Membr Biol 22:369–384CrossRefPubMedCentralGoogle Scholar
  11. Peña A, Cinco G, Gómez-Puyou A, Tuena de Gómez M (1972) Effects of the pH of the incubation medium on glycolysis and respiration in Saccharomyces cerevisiae. Arch Biochem Biophys 153:413–425CrossRefPubMedCentralGoogle Scholar
  12. Peña A, Clemente SM, Borbolla M, Carrasco N, Uribe S (1980) Multiple interactions of ethidium bromide with yeast cells. Arch Biochem Biophys 201:420–428CrossRefPubMedCentralGoogle Scholar
  13. Peña A, Uribe S, Pardo JP, Borbolla M (1984) The use of a cyanine dye in measuring membrane potential in yeast. Arch Biochem Biophys 231:217–225CrossRefPubMedCentralGoogle Scholar
  14. Peña A, Sánchez NS, Calahorra M (2010) Estimation of the electric plasma membrane potential difference in yeast with fluorescent dyes: comparative study of methods. J Bioenerg Biomembr 42:419–432CrossRefPubMedCentralGoogle Scholar
  15. Peña A, Sánchez NS, Calahorra M (2013) Effects of chitosan on Candida albicans: Conditions for its antifungal activity. Biomed Res Int 2013:527549. doi: 10.1155/2013/527549 CrossRefPubMedPubMedCentralGoogle Scholar
  16. Plášek J, Gášková D, Lichtenberg-Fraté H, Ludwig J, Höfer M (2012) Monitoring of real changes of plasma membrane potential by diS-C(3)(3) fluorescence in yeast cell suspensions. J Bioenerg Biomembr 44:559–569CrossRefPubMedCentralGoogle Scholar
  17. Sánchez NS, Arreguín R, Calahorra M, Peña A (2008) Effects of salts on aerobic metabolism of Debaryomyces hansenii. FEMS Yeast Res 8:1303–1312CrossRefPubMedCentralGoogle Scholar
  18. Sigler K, Höfer M (1991) Activation of the plasma membrane H(+)-ATPase of Saccharomyces cerevisiae by addition of hydrogen peroxide. Biochem Int 23:861–873PubMedPubMedCentralGoogle Scholar
  19. Vacata V, Kotyk A, Sigler K (1981) Membrane potentials in yeast cells measured by direct and indirect methods. Biochim Biophys Acta 643:265–268CrossRefPubMedCentralGoogle Scholar

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

Personalised recommendations