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Título del libro: Membrane Potential: An Overview
Título del capítulo: Estimations and Actual Measurements of the Plasma Membrane Electric Potential Difference in Yeast

Autores UNAM:
ANTONIO CALVIN PEÑA DIAZ; NORMA SILVIA SANCHEZ SANCHEZ; MARTHA EYDI RUSE CALAHORRA FUERTES;
Autores externos:

Idioma:

Año de publicación:
2019
Palabras clave:

fluorescence; plasma membrane potential; S. cerevisiae; thioflavin T; yeast


Resumen:

Estimation of the plasma membrane potential (PMP) in yeast by the fluorescence changes of various indicators have been studied by several groups for many years; with most variable results. The most used of these indicators have been DiSC3(3) and DiSC3(5). This contribution explores different dyes studied, methods and incubation media, as well as the different parameters analyzed. Particularly with DiSC3(3), from rather high to very low values of PMP have been calculated. Recently, we reported that fluorescence changes and accumulation of acridine yellow can be used to estimate and obtain actual values of the PMP in yeast. The experiments were performed with an old dye from a flask labeled ?acridine yellow? from a commercial source. However, NMR and mass spectrometry revealed that it was thioflavin T. With the pure dye, the experiments were repeated. Also the accumulation of the dye was measured to obtain real values of PMP, mainly based in permeabilizing the cells with chitosan in the absence or presence of an adequate concentration of KCl that allowed to correct the raw data obtained. Hence, more accurate values were obtained. Moreover, results of comparing this dye with others used so far, point to thioflavin T as the best one to follow by fluorescence and measure by its accumulation the plasma membrane potential in yeast. © 2019 by Nova Science Publishers, Inc.


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