Evaluation of mitochondrial membrane potential using a computerized device with a tetraphenylphosphonium-selective electrode

Mitochondrial membrane potential (Δψ m) plays important roles in the normal function of cells and in pathobiochemical situations. The application of ion-selective electrodes for the measurement of Δψ m is important for studying normal biological reactions and pathways and mitochondrial diseases. We...

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Bibliographic Details
Published in:Analytical biochemistry 2006-06, Vol.353 (1), p.37-42
Main Authors: Labajova, Anna, Vojtiskova, Alena, Krivakova, Pavla, Kofranek, Jiri, Drahota, Zdenek, Houstek, Josef
Format: Article
Language:English
Subjects:
Animals
Computer Peripherals
Computer Systems
Cultured cells
Electrophysiology - instrumentation
Electrophysiology - methods
Hepatocytes
Humans
Indicators and Reagents - chemistry
Ion-Selective Electrodes
Male
Membrane potential
Membrane Potentials - physiology
Mitochondria
Mitochondria, Liver - metabolism
Mitochondrial Membranes - metabolism
Onium Compounds - chemistry
Organophosphorus Compounds - chemistry
Rats
Rats, Wistar
Reproducibility of Results
TPP-selective electrode
Tumor Cells, Cultured
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Summary:Mitochondrial membrane potential (Δψ m) plays important roles in the normal function of cells and in pathobiochemical situations. The application of ion-selective electrodes for the measurement of Δψ m 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 Δψ 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 Δψ 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.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2006.03.032