Description
Abbreviation: ce
Reference: A Huetter 2004 Biochem J - SUIT-003
SUIT number: D009_ce1;ce2Omy;ce3U;ce4Rot;ce5Ama
O2k-Application: O2
- SUIT-category: ce
- SUIT protocol pattern: linear cell (ce) coupling control protocol 1ce;2ceOmy;3ceU
Template:SUIT-003 O2 O2 ce D009
References
Year | Reference | Organism | Tissue;cell | |
---|---|---|---|---|
MiPNet08.09 CellRespiration | 2016-08-11 | High-resolution respirometry and coupling-control protocol with living cells: ROUTINE, LEAK, ET-pathway, ROX. | Human | Blood cells Lymphocyte |
Gnaiger 2008 POS | 2008 | Gnaiger E (2008) Polarographic oxygen sensors, the oxygraph and high-resolution respirometry to assess mitochondrial function. In: Mitochondrial dysfunction in drug-induced toxicity (Dykens JA, Will Y, eds) John Wiley & Sons, Inc, Hoboken, NJ:327-52. | Mouse | Blood cells |
Huetter 2004 Biochem J | 2004 | Hütter E, Renner K, Pfister G, Stöckl P, Jansen-Dürr P, Gnaiger E (2004) Senescence-associated changes in respiration and oxidative phosphorylation in primary human fibroblasts. https://doi.org/10.1042/BJ20040095 | Human | Fibroblast |
R;L;E format_ROX
- The ceCCP diagram may show the respiratory states directly (left) as an alternative to presenting the titration steps.
- Rot and Ama
- In this ceCCP, ROX is measured after titration of rotenone, antimycin A, or both in a single step. Validity of using rotenone only for inducing ROX in intact cells is evaluated by stepwise measurement of ROX after titration of rotenone (3Rot) followed by titration of animycin A (4ceAma).
ce1;ce2Omy;ce3U;ce4Rot;ce5Ama
- ce1;ce2Omy;ce3U;ce4Rot;ce5Ama
- In this ceCCP, ROX is measured after stepwise titration of rotenone (ce4Rot) and antimycin A (ce5Ama). In many intact cells ROX measured after titration of rotenone (ce4Rot) is not or only slightly further inhibited by subsequent titration of animycin A (ce5Ama).
- Respiration in senescent human primary fibroblasts (0.2×106 cells/ml). Arrows show steps in the titration regime of the coupling control protocol, inducing the following respiratory states.
- 1ce: ROUTINE (R; routine state in cell-culture medium) in intact cells (ce);
- 2ceOmy: LEAK (L; inhibition of ATPsynthase by 1 μg/ml oligomycin);
- 3ceU: ET-pathway (E; maximal stimulation by uncoupling of oxidative phosphorylation in four subsequent titrations of the uncoupler, U, FCCP (2.5–4 μM final concentration);
- 4ceRot: ROX (rotenone, Rot; residual oxygen consumption).
- 5ceAma: ROX (antimycin A, Ama; residual oxygen consumption). Modified after Huetter_2004_Biochem J.
- Respiration in senescent human primary fibroblasts (0.2×106 cells/ml). Arrows show steps in the titration regime of the coupling control protocol, inducing the following respiratory states.
- Hütter E, Renner K, Pfister G, Stöckl P, Jansen-Dürr P, Gnaiger E (2004) Senescence-associated changes in respiration and oxidative phosphorylation in primary human fibroblasts. Biochem J 380:919-28. - »Bioblast link«
MitoPedia concepts:
SUIT protocol,
SUIT A,
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MitoPedia methods:
Respirometry