Labajova 2006 Anal Biochem

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.

» 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: MiParea: Respiration, Instruments;methods  Pathology: Cancer 

Organism: Rat  Tissue;cell: Liver  Preparation: Permeabilized cells, Isolated mitochondria 

Regulation: mt-Membrane potential 

HRR: Oxygraph-2k, TPP