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首页> 外文期刊>The Biochemical Journal >KINETIC STUDY OF THE PLASMA-MEMBRANE POTENTIAL IN PROCYCLIC AND BLOODSTREAM FORMS OF TRYPANOSOMA BRUCEI BRUCEI USING THE FLUORESCENT PROBE BISOXONOL
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KINETIC STUDY OF THE PLASMA-MEMBRANE POTENTIAL IN PROCYCLIC AND BLOODSTREAM FORMS OF TRYPANOSOMA BRUCEI BRUCEI USING THE FLUORESCENT PROBE BISOXONOL

机译：利用荧光探针比索诺尔对锥虫轮虫和循环血中血浆膜电位的运动学研究

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摘要

The characteristics of the plasma-membrane potential of procyclic and bloodstream forms of Trypanosoma brucei brucei (cultured cells) were investigated using the fluorescent anionic probe bisoxonol. Observation of a stable and representative plasma-membrane potential in the resting state required careful washing, centrifugation and maintenance of the cells at room temperature before measurement. Bloodstream forms were more prone to depolarization during washing at 4 degrees C than procyclic cells, The higher fluorescence observed in the presence of long slender cells than in the presence of procyclic cells shows that the plasma-membrane potential is more negative in the insect form. Healthy dilute cells can sustain their plasma-membrane potential for hours in the presence of external glucose. The presence of a high K+ concentration in the medium did not promote by itself the depolarization of either type of cell. Study of bisoxonol fluorescence as a function of time allowed us to follow the kinetics of the action of metabolic inhibitors in the presence of various ions. o-Vanadate (1 mM) was found to depolarize bloodstream-form cells rapidly but only in a phosphate-free NaCl buffer. Omeprazole and strophanthidin also specifically depolarized bloodstream-form trypanosomes. However, NN'-dicyclohexylcarbodi-imide depolarized both types of cell, but more rapidly for bloodstream-form cells. Bloodstream-form trypanosomes appear to use mainly a vanadate-sensitive Na+ pump to maintain their Na+-diffusion gradient. However, most of the ATPase inhibitors tested had little or no effect on the plasma-membrane potential of procyclics suggesting that this form of trypanosome may rely on several regulation mechanisms. [References: 32]
机译：布鲁氏锥虫（培养的细胞）的前周期和血流形式的血浆膜电位特性用荧光阴离子探针双氧嘧啶进行了研究。在静止状态下观察到稳定且有代表性的血浆膜电位需要在测量前仔细清洗，离心并保持细胞在室温下。在4°C的洗涤过程中，血流形式比周期细胞更易于去极化。在存在长而细长的细胞的情况下，所观察到的荧光要比在周期细胞中的更高，这表明在昆虫形式下，血浆膜电位更负。在存在外部葡萄糖的情况下，健康的稀释细胞可以维持数小时的血浆膜电位。培养基中高K +浓度的存在本身不会促进任何一种细胞的去极化。对比索诺尔荧光随时间变化的研究使我们能够在存在各种离子的情况下跟踪代谢抑制剂作用的动力学。发现邻钒酸盐（1 mM）可快速使血流形式的细胞去极化，但只能在无磷酸盐的NaCl缓冲液中进行。奥美拉唑和stophanthidin也特别去极化了血流形式的锥虫。然而，NN'-二环己基碳二亚胺使两种类型的细胞去极化，但对于血流形式的细胞则更快。血流形式的锥虫似乎主要使用对钒酸盐敏感的Na +泵来维持其Na +扩散梯度。但是，测试的大多数ATPase抑制剂对脯氨酸的血浆膜电位几乎没有影响，甚至没有影响，表明这种形式的锥虫可能依赖于几种调节机制。 [参考：32]

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