Relation between mitochondrial membrane potential and ROS formation

Methods Mol Biol. 2012;810:183-205. doi: 10.1007/978-1-61779-382-0_12.

Abstract

Mitochondria are considered as the main source of reactive oxygen species (ROS) in the cell. For this reason, they have been recognized as a source of various pathological conditions as well as aging. Chronic increase in the rate of ROS production is responsible for the accumulation of ROS-associated damages in DNA, proteins, and lipids, and may result in progressive cell dysfunctions and, in a consequence, apoptosis, increasing the overall probability of an organism's pathological conditions. The superoxide anion is the main undesired by-product of mitochondrial oxidative phosphorylation. Its production is triggered by a leak of electrons from the mitochondrial respiratory chain and the reaction of these electrons with O(2). Superoxide dismutase (MnSOD, SOD2) from the mitochondrial matrix as well as superoxide dismutase (Cu/ZnSOD, SOD1) present in small amounts in the mitochondrial intramembrane space, convert superoxide anion to hydrogen peroxide, which can be then converted by catalase to harmless H(2)O. In this chapter, we describe a relation between mitochondrial membrane potential and the rate of ROS formation. We present different methods applicable for isolated mitochondria or intact cells. We also present experiments demonstrating that a magnitude and a direction (increase or decrease) of a change in mitochondrial ROS production depends on the metabolic state of this organelle.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Benzimidazoles / metabolism
  • Brain / metabolism
  • Calcium / metabolism
  • Carbocyanines / metabolism
  • Carcinoma, Ehrlich Tumor / metabolism
  • Cell Fractionation / methods
  • Cell Line, Tumor
  • Electron Transport
  • Fibroblasts / metabolism
  • HeLa Cells
  • Humans
  • Hydrogen Peroxide / metabolism
  • Membrane Potential, Mitochondrial*
  • Mice
  • Microscopy, Confocal
  • Mitochondria / metabolism*
  • Oxygen Consumption
  • Phenazines / metabolism
  • Reactive Oxygen Species / metabolism*
  • Superoxides / metabolism

Substances

  • Benzimidazoles
  • Carbocyanines
  • Phenazines
  • Reactive Oxygen Species
  • Superoxides
  • 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine
  • Hydrogen Peroxide
  • Calcium
  • safranine T