Journal
JOURNAL OF BIOENERGETICS AND BIOMEMBRANES
Volume 42, Issue 5, Pages 419-432Publisher
SPRINGER/PLENUM PUBLISHERS
DOI: 10.1007/s10863-010-9311-x
Keywords
Yeast; Membrane potential indicators; Ion transport
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Funding
- Consejo Nacional de Ciencia y Tecnologia de Mexico [46521]
- Direccion de Asuntos del Personal Academico of this University [IN200307]
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Different methods to estimate the plasma membrane potential difference (PMP) of yeast cells with fluorescent monitors were compared. The validity of the methods was tested by the fluorescence difference with or without glucose, and its decrease by the addition of 10 mM KCl. Low CaCl2 concentrations avoid binding of the dye to the cell surface, and low CCCP concentrations avoid its accumulation by mitochondria. Lower concentrations of Ba2+ produce a similar effect as Ca2+, without producing the fluorescence changes derived from its transport. Fluorescence changes without considering binding of the dyes to the cells and accumulation by mitochondria are overshadowed by their distribution between this organelle and the cytoplasm. Other factors, such as yeast starvation, dye used, parameters of the fluorescence changes, as well as buffers and incubation times were analyzed. An additional approach to measure the actual or relative values of PMP, determining the accumulation of the dye, is presented.
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