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  O2 affects mitochondrial functionality ex vivo.

Nanadikara, M. S., Vergel Leon, A. M., Borowik, S., Hillemann, A., Zieseniss, A., Belousov, V. V., et al. (2019). O2 affects mitochondrial functionality ex vivo. Redox Biology, 22: 101152. doi:10.1016/j.redox.2019.101152.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0003-1CC1-4 Version Permalink: http://hdl.handle.net/21.11116/0000-0003-1CCC-9
Genre: Journal Article

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 Creators:
Nanadikara, M. S., Author
Vergel Leon, A. M., Author
Borowik, S., Author
Hillemann, A., Author
Zieseniss, A., Author
Belousov, V. V., Author
Bogeski, I., Author
Rehling, P.1, Author              
Dudek, J., Author
Katschinski, D. M., Author
Affiliations:
1Max Planck Fellow Peter Rehling, ou_1298545              

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Free keywords: Glutathione redox potential; Grx1-roGFP; Hypoxia; Mitochondrial matrix
 Abstract: Mitochondria have originated in eukaryotic cells by endosymbiosis of a specialized prokaryote approximately 2 billion years ago. They are essential for normal cell function by providing energy through their role in oxidizing carbon substrates. Glutathione (GSH) is a major thiol-disulfide redox buffer of the cell including the mitochondrial matrix and intermembrane space. We have generated cardiomyocyte-specific Grx1-roGFP2 GSH redox potential (EGSH) biosensor mice in the past, in which the sensor is targeted to the mitochondrial matrix. Using this mouse model a distinct EGSH of the mitochondrial matrix (-278.9 ± 0.4 mV) in isolated cardiomyocytes is observed. When analyzing the EGSH in isolated mitochondria from the transgenic hearts, however, the EGSH in the mitochondrial matrix is significantly oxidized (-247.7 ± 8.7 mV). This is prevented by adding N-Ethylmaleimide during the mitochondria isolation procedure, which precludes disulfide bond formation. A similar reducing effect is observed by isolating mitochondria in hypoxic (0.1-3% O2) conditions that mimics mitochondrial pO2 levels in cellulo. The reduced EGSH is accompanied by lower ROS production, reduced complex III activity but increased ATP levels produced at baseline and after stimulation with succinate/ADP. Altogether, we demonstrate that oxygenation is an essential factor that needs to be considered when analyzing mitochondrial function ex vivo.

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Language(s): eng - English
 Dates: 2019-02-232019-04
 Publication Status: Published in print
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 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1016/j.redox.2019.101152
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Title: Redox Biology
Source Genre: Journal
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Pages: 8 Volume / Issue: 22 Sequence Number: 101152 Start / End Page: - Identifier: -