A method to assess the mitochondrial respiratory capacity of complexes I and II from frozen tissue using the Oroboros O2k-FluoRespirometer

Ebanks, Brad; Kwiecinska, Pola; Moisoi, Nicoleta; Chakrabarti, Lisa

Abstract

High-resolution respirometry methods allow for the assessment of oxygen consumption by the electron transfer systems within cells, tissue samples, and isolated mitochondrial preparations. As mitochondrial integrity is compromised by the process of cryopreservation, these methods have been limited to fresh samples. Here we present a simple method to assess the activity of mitochondria respiratory complexes I and II in previously cryopreserved murine skeletal muscle tissue homogenates, as well as previously frozen D. melanogaster, as a function of oxygen consumption.

Citation

Ebanks, B., Kwiecinska, P., Moisoi, N., & Chakrabarti, L. (2023). A method to assess the mitochondrial respiratory capacity of complexes I and II from frozen tissue using the Oroboros O2k-FluoRespirometer. PLoS ONE, 18(7), Article e0276147. https://doi.org/10.1371/journal.pone.0276147

• Journal Article Type: Article
• Acceptance Date: May 30, 2023
• Online Publication Date: Jul 24, 2023
• Publication Date: Jul 24, 2023
• Deposit Date: Aug 18, 2023
• Publicly Available Date: Aug 21, 2023
• Journal: PLoS ONE
• Electronic ISSN: 1932-6203
• Publisher: Public Library of Science
• Peer Reviewed: Peer Reviewed
• Volume: 18
• Issue: 7
• Article Number: e0276147
• DOI: https://doi.org/10.1371/journal.pone.0276147
• Keywords: Drosophila melanogaster; Mitochondria; Oxygen consumption; Skeletal muscles; Oxygen; Electron transfer; Respirometry; Mitochondrial membrane
• Public URL: https://nottingham-repository.worktribe.com/output/23472699
• Publisher URL: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0276147