Labajova 2006 Anal Biochem: Difference between revisions
No edit summary |
No edit summary |
||
Line 7: | Line 7: | ||
|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, | ||
|mipnetlab=CZ_Hradec Kralove_Cervinkova Z, CZ_Prague_Houstek J | |mipnetlab=CZ_Hradec Kralove_Cervinkova Z, CZ_Prague_Houstek J, CZ Prague Bioenergetics | ||
|discipline=Biomedicine | |discipline=Biomedicine | ||
}} | }} | ||
Line 16: | Line 16: | ||
|tissues=Hepatocyte; Liver | |tissues=Hepatocyte; Liver | ||
|preparations=Permeabilized cells, Isolated Mitochondria | |preparations=Permeabilized cells, Isolated Mitochondria | ||
|additional=Instrumental and methodological aspects | |additional=Instrumental and methodological aspects | ||
|discipline=Biomedicine | |discipline=Biomedicine | ||
}} | }} |
Revision as of 13:47, 7 December 2012
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) Analyt 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, CZ Prague Bioenergetics
Labels:
Stress:Cancer; Apoptosis; Cytochrome c"Cancer; Apoptosis; Cytochrome c" is not in the list (Cell death, Cryopreservation, Ischemia-reperfusion, Permeability transition, Oxidative stress;RONS, Temperature, Hypoxia, Mitochondrial disease) of allowed values for the "Stress" property. Organism: Rat Tissue;cell: Hepatocyte; Liver"Hepatocyte; Liver" is not in the list (Heart, Skeletal muscle, Nervous system, Liver, Kidney, Lung;gill, Islet cell;pancreas;thymus, Endothelial;epithelial;mesothelial cell, Blood cells, Fat, ...) of allowed values for the "Tissue and cell" property. Preparation: Permeabilized cells, Isolated Mitochondria"Isolated Mitochondria" is not in the list (Intact organism, Intact organ, Permeabilized cells, Permeabilized tissue, Homogenate, Isolated mitochondria, SMP, Chloroplasts, Enzyme, Oxidase;biochemical oxidation, ...) of allowed values for the "Preparation" property.
HRR: Oxygraph-2k, TPP
Instrumental and methodological aspects