Labajova 2006 Anal Biochem: Difference between revisions
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|authors=Labajova A, Vojtiskova A, Krivakova P, Kofranek J, Drahota Z, Houstek J | |authors=Labajova A, Vojtiskova A, Krivakova P, Kofranek J, Drahota Z, Houstek J | ||
|year=2006 | |year=2006 | ||
|journal= | |journal=Anal Biochem | ||
|abstract=Mitochondrial membrane potential (ฮฯm) plays important roles in the normal function of cells and in pathobiochemical situations. The application of ion-selective electrodes for the measurement of ฮฯmย 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 ฮฯm, 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 ฮฯm (as changes in TPP+ย 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. | |abstract=Mitochondrial membrane potential (ฮฯm) plays important roles in the normal function of cells and in pathobiochemical situations. The application of ion-selective electrodes for the measurement of ฮฯmย 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 ฮฯm, 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 ฮฯm (as changes in TPP+ย 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. | ||
|keywords=Membrane potential, TPP-selective electrode, Mitochondria, Hepatocytes, Cultured cells | |keywords=Membrane potential, TPP-selective electrode, Mitochondria, Hepatocytes, Cultured cells |
Revision as of 23:12, 25 December 2015
Labajova A, Vojtiskova A, Krivakova P, Kofranek J, Drahota Z, Houstek J (2006) Evaluation of mitochondrial membrane potential using a computerized device with a tetraphenylphosphonium-selective electrode. Anal Biochem 353:37-42. |
Labajova A, Vojtiskova A, Krivakova P, Kofranek J, Drahota Z, Houstek J (2006) Anal Biochem
Abstract: Mitochondrial membrane potential (ฮฯm) plays important roles in the normal function of cells and in pathobiochemical situations. The application of ion-selective electrodes for the measurement of ฮฯm 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 ฮฯm, 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 ฮฯm (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. โข Keywords: Membrane potential, TPP-selective electrode, Mitochondria, Hepatocytes, Cultured cells
โข O2k-Network Lab: CZ Hradec Kralove Cervinkova Z, CZ Prague Houstek J
Labels: MiParea: Respiration, Instruments;methods
Pathology: Cancer
Organism: Rat Tissue;cell: Liver Preparation: Permeabilized cells, Isolated mitochondria
Regulation: mt-Membrane potential
HRR: Oxygraph-2k, TPP