Faculty of Mathematics and Physics, Charles University in Prague, Prague, Czech Republic,
J Bioenerg Biomembr. 2013 Dec;45(6):561-8. doi: 10.1007/s10863-013-9528-6. Epub 2013 Sep 20.
Recently we introduced a fluorescent probe technique that makes possible to convert changes of equilibrium fluorescence spectra of 3,3'-dipropylthiadicarbocyanine, diS-C3(3), measured in yeast cell suspensions under defined conditions into underlying membrane potential differences, scaled in millivolts (Plasek et al. in J Bioenerg Biomembr 44: 559-569, 2012). The results presented in this paper disclose measurements of real early changes of plasma membrane potential induced by the increase of extracellular K(+), Na(+) and H(+) concentration in S. cerevisiae with and without added glucose as energy source. Whereas the wild type and the ∆tok1 mutant cells exhibited similar depolarization curves, mutant cells lacking the two Trk1,2 potassium transporters revealed a significantly decreased membrane depolarization by K(+), particularly at lower extracellular potassium concentration [K(+)]out. In the absence of external energy source plasma membrane depolarization by K(+) was almost linear. In the presence of glucose the depolarization curves exhibited an exponential character with increasing [K(+)]out. The plasma membrane depolarization by Na(+) was independent from the presence of Trk1,2 transporters. Contrary to K(+), Na(+) depolarized the plasma membrane stronger in the presence of glucose than in its absence. The pH induced depolarization exhibited a fairly linear relationship between the membrane potential and the pHo of cell suspensions, both in the wild type and the Δtrk1,2 mutant strains, when cells were energized by glucose. In the absence of glucose the depolarization curves showed a biphasic character with enhanced depolarization at lower pHo values.
最近,我们引入了一种荧光探针技术,使得将 3,3'-二丙基噻二碳菁二磺酸钠(diS-C3(3))在酵母悬浮液中在规定条件下的平衡荧光光谱变化转换为潜在的膜电位差成为可能,其以毫伏为单位进行缩放(Plasek 等人,J Bioenerg Biomembr 44: 559-569, 2012)。本文介绍的结果揭示了测量由于细胞外 K(+)、Na(+)和 H(+)浓度的增加而引起的真核生物细胞膜电位的早期变化,这些测量是在有和没有添加葡萄糖作为能源的情况下进行的。野生型和 ∆tok1 突变细胞表现出相似的去极化曲线,而缺乏两个 Trk1、2 钾转运体的突变细胞显示出由 K(+)引起的膜去极化显著降低,特别是在较低的细胞外钾浓度[K(+)]out 时。在没有外部能源的情况下,K(+)引起的质膜去极化几乎呈线性。在存在葡萄糖的情况下,去极化曲线呈指数特征,随着[K(+)]out 的增加而增加。Na(+)引起的质膜去极化与 Trk1、2 转运体的存在无关。与 K(+)相反,Na(+)在有葡萄糖的情况下比在没有葡萄糖的情况下更强烈地去极化质膜。在葡萄糖为细胞供能时,pH 诱导的去极化与细胞悬浮液的膜电位和 pHo 之间存在相当线性的关系,无论是在野生型还是在Δtrk1、2 突变株中。在没有葡萄糖的情况下,去极化曲线呈现出双相特征,在较低的 pHo 值下出现增强的去极化。