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Evaluation of mitochondrial membrane potential measured by the computerized device using tetraphenylphosphonium‐selective electrode
Author(s) -
Labajova Anna,
Drahota Zdenek,
Vojtiskova Alena,
Vrbacky Marek,
Krivakova Pavla,
Cerkasov Jiri,
Kofranek Jiri
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a525-d
Subject(s) - membrane potential , mitochondrion , membrane , chemistry , respiratory chain , biophysics , mitochondrial matrix , electrode , hela , biochemistry , cell , microbiology and biotechnology , biology , enzyme , cytosol
Mitochondria play an essential role in cell energy transformation. One of the most important indicators of the mitochondrial function is the mitochondrial membrane potential. A device for computerized monitoring of mitochondrial membrane potential was constructed and preparation of tetraphenylphosphonium (TPP + )‐selective electrode, construction of a measuring chamber and electrodes were optimized. The method is based on the accumulation of TPP + in the mitochondrial matrix according to the membrane potential and it allows to calculate the membrane potential of isolated mitochondria in the absolute scale of millivolts. Our improved device was successfully used for the membrane potential measurement of isolated mitochondria and mitochondria of permeabilized cells (hepatocytes, cybrides, HeLa G and BSC‐40). Moreover, the influence of substrates, inhibitors and uncouplers of respiratory chain was studied. The optimized device we designed is suitable for characterization of mitochondrial diseases associated with defects of respiratory chain enzyme complexes, which are often reflected in changes of the mitochondrial membrane potential. This work was supported by Ministry of Education of the Czech Republic grant MSM‐111100008, MSMT 394 and MSM‐0021620806.