| Chapter title |
Assaying Mitochondrial Respiration as an Indicator of Cellular Metabolism and Fitness
|
|---|---|
| Chapter number | 7 |
| Book title |
Cell Viability Assays
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-6960-9_7 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6959-3, 978-1-4939-6960-9
|
| Authors |
Natalia Smolina, Joseph Bruton, Anna Kostareva, Thomas Sejersen, Smolina, Natalia, Bruton, Joseph, Kostareva, Anna, Sejersen, Thomas |
| Editors |
Daniel F. Gilbert, Oliver Friedrich |
| Abstract |
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. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 1 | 50% |
| Unknown | 1 | 50% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 2 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 76 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Master | 17 | 22% |
| Student > Bachelor | 11 | 14% |
| Student > Ph. D. Student | 10 | 13% |
| Researcher | 4 | 5% |
| Professor | 3 | 4% |
| Other | 7 | 9% |
| Unknown | 24 | 32% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 17 | 22% |
| Immunology and Microbiology | 6 | 8% |
| Medicine and Dentistry | 6 | 8% |
| Agricultural and Biological Sciences | 4 | 5% |
| Pharmacology, Toxicology and Pharmaceutical Science | 3 | 4% |
| Other | 14 | 18% |
| Unknown | 26 | 34% |