Evaluation of mitochondrial membrane potential using a computerized device with a tetraphenylphosphonium-selective electrode
Evaluation of mitochondrial membrane potential using a computerized device with a tetraphenylphosphonium-selective electrode
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DOI：
10.1016/j.ab.2006.03.032
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发表时间：
2006-06-01
影响因子：
2.9
通讯作者：
Houstek, J
中科院分区：
文献类型：
--
作者：
Labajova, A;Vojtiskova, A;Houstek, J
线粒体膜电位（δ psi(m)）在细胞的正常功能和病理生化情况下起着重要作用。应用离子选择电极测量A，对于研究正常的生物反应和途径以及线粒体疾病具有重要意义。我们构建并优化了用于实时监测δ psi(m)的计算机设备，其中包括对四苯基磷（TPP+）选择膜的修饰，以提高TPP+选择电极的可重复性。应用MATLAB软件提高了系统的灵敏度。我们测试了改进后的装置，用于分离线粒体的膜电位测量（以毫伏为绝对尺度）。此外，我们评估了添加底物后洋地黄苷渗透细胞（肝细胞、对照线粒体胞体、HeLa G和BSC-40）的δ psi(m) （TPP+浓度的变化）的相对变化。抑制剂。和呼吸链的解耦器。我们的系统可以成功地用于线粒体生物能量调节的许多方面的研究，并作为线粒体氧化磷酸化疾病的诊断工具。(c) 2006爱思唯尔公司版权所有。
Mitochondrial membrane potential (Delta psi(m)) plays important roles in the normal function of cells and in pathobiochemical situations. The application of ion-selective electrodes for the measurement of A,, is important for studying normal biological reactions and pathways and mitochondrial diseases. We constructed and optimized a computerized device for real-time monitoring of the Delta psi(m), which included modification of tetraphenylphosphonium (TPP+)-selective membrane that improved reproducibility of the TPP+-selective electrode. Application of MATLAB software increased the sensitivity of the system. We tested our improved device for membrane potential measurements of isolated mitochondria (in absolute scale of millivolts). In addition, we assessed relative changes of Delta psi(m) (as changes in TPP+ concentration) of digitonin-permeabilized cells (hepatocytes, control transmitochondrial cybrids, HeLa G and BSC-40) after addition of substrates. inhibitors. and uncoupler of respiratory chain. Our system can be Successfully used for Studies of many aspects of the regulation of mitochondrial bioenergetics and as a diagnostic tool for mitochondrial oxidative phosphorylation disorders. (c) 2006 Elsevier Inc. All rights reserved.