SUIT-014 O2 pfi D042: Difference between revisions
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::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS(PGM) | ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS(PGM) | ||
::: '''[[SUIT protocol pattern]]:''' diametral 1GM;2D;3P;4S;5U;6Rot | ::: '''[[SUIT protocol pattern]]:''' diametral 1GM;2D;3P;4S;5U;6Rot- | ||
__TOC__ | __TOC__ |
Revision as of 11:57, 26 February 2019
Description
Abbreviation: NS(PGM)
Reference: A
SUIT number: D042_1GM;2D;3P;4S;5U;6Rot;7Ama
O2k-Application: O2
- SUIT-category: NS(PGM)
- SUIT protocol pattern: diametral 1GM;2D;3P;4S;5U;6Rot-
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
1GM | GML(n) | N | CI | 1GM
|
2D | GMP | N | CI | 1GM;2D
|
2c | GMcP | N | CI | 1GM;2D;2c
|
3P | PGMP | N | CI | 1GM;2D;2c;3P
|
4S | PGMSP | NS | CI&II | 1GM;2D;2c;3P;4S
|
5U | PGMSE | NS | CI&II | 1GM;2D;2c;3P;4S;5U
|
6Rot | SE | S | CII | 1GM;2D;2c;3P;4S;5U;6Rot
|
7Ama | ROX | 1GM;2D;2c;3P;4S;5U;6Rot;7Ama
|
Step | Respiratory state | Pathway control | ET-Complex | Comment |
---|---|---|---|---|
## AsTm | AsTmE | CIV | CIV | |
## Azd | CHB |
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- Coupling control
- Pathway control
- ยป Electron transfer pathway
- ยป Fatty acid oxidation pathway control state, F
- ยป NADH electron transfer-pathway state, N
- ยป Succinate pathway control state, S
- ยป NS-pathway control state, NS
- ยป Glycerophosphate pathway control state, Gp
- ยป Complex IV single step, CIV
- ยป Anaplerotic pathway control state
- Pathway control
- Main fuel substrates
- ยป Glutamate, G
- ยป Glycerophosphate, Gp
- ยป Malate, M
- ยป Octanoylcarnitine, Oct
- ยป Pyruvate, P
- ยป Succinate, S
- Main fuel substrates
- Glossary
Strengths and limitations
- + NS-OXPHOS capacity provides a physiologically relevant estimate of maximum mitochondrial respiratory capacity.
- + The presence of PGM and S establishes fully operative TCA cycle activity.
- + This protocol allows to analyse the convergence of pathways at the Q-junction (N, NS, S).
- + Mitochondrial external membrane integrity can be measured with the addition of cytochrome c. Application of the cytochrome c test early in the protocol ensures comparability of all states in case of any effect of c.
- - It was observed that addition of P (5 mM) after GM inhibited respiration in horse skeletal muscle fibres (Votion et al 2012), which was not the case when P was titrated in steps of 1 mM.
- - When evaluating the additive effect of the N- and S-pathway, it has to be considered that NSP- and NSE-capacities can only be compared with NP- and SE-capacities. This is not a problem when NSP = NSE (Gnaiger 2009). Otherwise, it may be assumed that SP = SE (Votion et al 2012), such that NSP can be compared with NP + SP. SUIT-004 should be chosen for the additive effect in the ET-state.
- - Rox may be lower in substrate states earlier in the SUIT protocol. Therefore, this Rox measurement is frequently taken as a methodological control rather than as the final basis of Rox correction of mitochondrial respiration (mt).
- - Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler.
- - CIV activity is not measured, to save experimental time.
Compare SUIT protocols
References
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry