A New Strategy to Preserve and Assess Oxygen Consumption in Murine Tissues.
Jerome KluzaVictoriane PeugnetBlanche DaunouWilliam LaineGwenola KervoazeGaëlle RémyAnne LoyensPatrice MaboudouQuentin FovezCorinne GrangetteIsabelle WolowczukPhilippe GossetGuillaume GarçonPhilippe MarchettiFlorence PinetMuriel PichavantEmilie Dubois-DeruyPublished in: International journal of molecular sciences (2021)
Mitochondrial dysfunctions are implicated in several pathologies, such as metabolic, cardiovascular, respiratory, and neurological diseases, as well as in cancer and aging. These metabolic alterations are usually assessed in human or murine samples by mitochondrial respiratory chain enzymatic assays, by measuring the oxygen consumption of intact mitochondria isolated from tissues, or from cells obtained after physical or enzymatic disruption of the tissues. However, these methodologies do not maintain tissue multicellular organization and cell-cell interactions, known to influence mitochondrial metabolism. Here, we develop an optimal model to measure mitochondrial oxygen consumption in heart and lung tissue samples using the XF24 Extracellular Flux Analyzer (Seahorse) and discuss the advantages and limitations of this technological approach. Our results demonstrate that tissue organization, as well as mitochondrial ultrastructure and respiratory function, are preserved in heart and lung tissues freshly processed or after overnight conservation at 4 °C. Using this method, we confirmed the repeatedly reported obesity-associated mitochondrial dysfunction in the heart and extended it to the lungs. We set up and validated a new strategy to optimally assess mitochondrial function in murine tissues. As such, this method is of great potential interest for monitoring mitochondrial function in cohort samples.
Keyphrases
- oxidative stress
- gene expression
- heart failure
- single cell
- induced apoptosis
- type diabetes
- endothelial cells
- cell therapy
- metabolic syndrome
- insulin resistance
- mental health
- stem cells
- nitric oxide
- adipose tissue
- papillary thyroid
- cell cycle arrest
- skeletal muscle
- signaling pathway
- brain injury
- human health
- induced pluripotent stem cells
- subarachnoid hemorrhage
- pi k akt