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Difference between revisions of "Labajova 2006 Anal Biochem"

From Bioblast
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|journal=Analyt. Biochem.
|journal=Analyt. Biochem.
|abstract=Mitochondrial membrane potential (Δψm) plays important roles in the normal function of cells and in pathobiochemical situations. The application of ion-selective electrodes for the measurement of Δψm  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 Δψm, 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 Δψm (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.
|abstract=Mitochondrial membrane potential (Δψm) plays important roles in the normal function of cells and in pathobiochemical situations. The application of ion-selective electrodes for the measurement of Δψm  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 Δψm, 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 Δψm (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.
|keywords=Membrane potential, TPP-selective electrode, Mitochondria, Hepatocytes, Cultured cells, Instrumental and methodological aspects
|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
|discipline=Biomedicine
|discipline=Biomedicine
Line 17: Line 17:
|preparations=Isolated Mitochondria, Permeabilized Cell or Tissue; Homogenate
|preparations=Isolated Mitochondria, Permeabilized Cell or Tissue; Homogenate
|topics=Coupling; Membrane Potential
|topics=Coupling; Membrane Potential
|additional=Instrumental and methodological aspects
|discipline=Biomedicine
|discipline=Biomedicine
}}
}}

Revision as of 10:55, 11 November 2011

Publications in the MiPMap
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. Analyt. Biochem. 353: 37-42.

» PMID: 16643832

Labajova A, Vojtiskova A, Krivakova P, Kofranek J, Drahota Z, Houstek J (2006) Analyt. Biochem.

Abstract: Mitochondrial membrane potential (Δψm) plays important roles in the normal function of cells and in pathobiochemical situations. The application of ion-selective electrodes for the measurement of Δψm 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 Δψm, 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 Δψm (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. Keywords: Membrane potential, TPP-selective electrode, Mitochondria, Hepatocytes, Cultured cells

O2k-Network Lab: CZ_Hradec Kralove_Cervinkova Z, CZ_Prague_Houstek J


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: 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., Permeabilized Cell or Tissue; Homogenate"Permeabilized Cell or Tissue; Homogenate" 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. 

Regulation: Coupling; Membrane Potential"Coupling; Membrane Potential" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property. 


HRR: Oxygraph-2k, TPP 

Instrumental and methodological aspects