Reactive oxygen species induce swelling and cytochrome c release but not transmembrane depolarization in isolated rat brain mitochondria.

In this study, we have used isolated brain mitochondria to investigate the effects of superoxide anions (O2-) on mitochondrial parameters related to apoptosis, such as swelling, potential, enzymatic activity, NAD(P)H, cytochrome c release, and caspase activity.Addition of the reactive oxygen species...

Full description

Bibliographic Details
Authors: Galindo Anaya, María Francisca, Jordán Bueso, Joaquín, González García, María del Carmen, Ceña Callejo, Valentín
Format: article
Publication Date:2003
Country:España
Institution:Universidad de Castilla-La Mancha
Repository:RUIdeRA. Repositorio Institucional de la UCLM
OAI Identifier:oai:ruidera.uclm.es:10578/33366
Online Access:https://hdl.handle.net/10578/33366
Access Level:Open access
Keyword:Apoptosis
Brain
Caspase
Excitotoxicity
Mitochondrion
ROS
Description
Summary:In this study, we have used isolated brain mitochondria to investigate the effects of superoxide anions (O2-) on mitochondrial parameters related to apoptosis, such as swelling, potential, enzymatic activity, NAD(P)H, cytochrome c release, and caspase activity.Addition of the reactive oxygen species (ROS) generator KO2 produced brain mitochondrial swelling, which was blocked by cyclosporin A (CSA), and which was Ca2+ independent.Calcium induced mitochondrial swelling only at high concentrations and in the presence of succinate. This correlated with the increase in O2- production detected with hydroethidine in mitochondrial preparations exposed to Ca2+ and the fact that ROS were required for Ca2+-induced mitochondrial swelling.Superoxide anions, but not Ca2+, decreased citrate synthase and dehydrogenase enzymatic activities and dropped total mitochondrial NAD(P)H levels.Calcium, but not O2-, triggered a rapid loss of mitochondrial potential. Calcium-induced ??m dissipation was inhibited by Ruthenium Red, but not by CSA.Calcium- and superoxide-induced mitochondrial swelling released cytochrome c and increased caspase activity from isolated mitochondria in a CS A-sensitive manner.In summary, superoxide potently triggers mitochondrial swelling and the release of proteins involved in activation of postmitochondrial apoptotic pathways in the absence of mitochondrial depolarization.