Assaying Mitochondrial Respiration as an Indicator of Cellular Metabolism and Fitness
| Autor: | Natalia, Smolina, Joseph, Bruton, Anna, Kostareva, Thomas, Sejersen |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
Electron Transport Complex I Satellite Cells Skeletal Muscle Cell Survival Uncoupling Agents Cell Respiration Muscle Fibers Skeletal Primary Cell Culture Antimycin A Oxidative Phosphorylation Mitochondria Electron Transport Complex III Mice Adenosine Triphosphate Oxygen Consumption Rotenone Proton Ionophores Animals Biological Assay Oligomycins |
| Zdroj: | Methods in molecular biology (Clifton, N.J.). 1601 |
| ISSN: | 1940-6029 |
| Popis: | Mitochondrial respiration is the most important generator of cellular energy under most circumstances. It is a process of energy conversion of substrates into ATP. The Seahorse equipment allows measuring oxygen consumption rate (OCR) in living cells and estimates key parameters of mitochondrial respiration in real-time mode. Through use of mitochondrial inhibitors, four key mitochondrial respiration parameters can be measured: basal, ATP production-linked, maximal, and proton leak-linked OCR. This approach requires application of mitochondrial inhibitors-oligomycin to block ATP synthase, FCCP-to make the inner mitochondrial membrane permeable for protons and allow maximum electron flux through the electron transport chain, and rotenone and antimycin A-to inhibit complexes I and III, respectively. This chapter describes the protocol of OCR assessment in the culture of primary myotubes obtained upon satellite cell fusion. |
| Databáze: | OpenAIRE |
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