Abstract
Mitochondria are considered 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 O2. 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, converts superoxide anion to hydrogen peroxide, which can be then converted by catalase to harmless H2O.
In the 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 depend on the metabolic state of this organelle.
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Acknowledgments
This work was supported by the Polish Ministry of Science and Higher Education grants N301 092 32/3407 and N N407 075 137 for MRW, JD, ML, and JS. JS was also supported by a PhD fellowship from the Foundation for Polish Science (FNP), UE, European Regional Development Fund, and Operational Programme “Innovative Economy.” PP and MB are supported by AIRC, Telethon (GGP09128); local funds from the University of Ferrara; the PRRIITT program of the Emilia-Romagna Region; the Italian Multiple Sclerosis Foundation (FISM Cod.2008/R/18); the Italian Ministry of Education, University and Research; and the Italian Ministry of Health.
Ethics
The studies with the use of human fibroblasts were carried out in accordance with the Declaration of Helsinki of the World Medical Association and were approved by the Committee on Bioethics at the Children’s Memorial Health Institute. Informed consent was obtained from the parents before any biopsy or molecular analysis was performed.
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Suski, J., Lebiedzinska, M., Bonora, M., Pinton, P., Duszynski, J., Wieckowski, M.R. (2018). Relation Between Mitochondrial Membrane Potential and ROS Formation. In: Palmeira, C., Moreno, A. (eds) Mitochondrial Bioenergetics. Methods in Molecular Biology, vol 1782. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7831-1_22
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DOI: https://doi.org/10.1007/978-1-4939-7831-1_22
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