TY - CHAP
T1 - Electrochemical detection of H2O2 formation in isolated mitochondria
AU - Rapino, Stefania
AU - Marcu, Raluca
AU - Paolucci, Francesco
AU - Giorgio, Marco
PY - 2013
Y1 - 2013
N2 - Mitochondrial respiration produces both complete and partially reduced oxygen species that are involved in physiological and pathological processes. Indeed, unspecific oxidative damage induced by excessive mitochondrial reactive oxygen species (ROS) plays a role in aging and several diseases, whereas low amounts of ROS act in physiological signaling processes. The exact molecular species, the rate, and the conditions of mitochondrial ROS release are not clearly evaluable by current methods based on oxidation sensitive markers. Recently, electrochemical analysis of biological samples has improved. Following latest methodology, we implemented a novel electrochemical assay for the investigation of aerobic metabolism in isolated mitochondria through simultaneous measurement of O2 consumption and H2O 2 production. Our experiments confirm active H2O 2 production by respiring mouse liver mitochondria and show that ATP synthase activation increases the rate of H2O2, suggesting that state 3 mitochondria might induce the cell through oxidative signals.
AB - Mitochondrial respiration produces both complete and partially reduced oxygen species that are involved in physiological and pathological processes. Indeed, unspecific oxidative damage induced by excessive mitochondrial reactive oxygen species (ROS) plays a role in aging and several diseases, whereas low amounts of ROS act in physiological signaling processes. The exact molecular species, the rate, and the conditions of mitochondrial ROS release are not clearly evaluable by current methods based on oxidation sensitive markers. Recently, electrochemical analysis of biological samples has improved. Following latest methodology, we implemented a novel electrochemical assay for the investigation of aerobic metabolism in isolated mitochondria through simultaneous measurement of O2 consumption and H2O 2 production. Our experiments confirm active H2O 2 production by respiring mouse liver mitochondria and show that ATP synthase activation increases the rate of H2O2, suggesting that state 3 mitochondria might induce the cell through oxidative signals.
KW - Electrochemistry
KW - Hydrogen peroxide
KW - Mitochondria
KW - Respiratory chain
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U2 - 10.1016/B978-0-12-405883-5.00007-7
DO - 10.1016/B978-0-12-405883-5.00007-7
M3 - Chapter
C2 - 23791097
AN - SCOPUS:84879759282
SN - 9780124058835
VL - 526
T3 - Methods in Enzymology
SP - 123
EP - 134
BT - Methods in Enzymology
ER -