SUIT-011 O2 pfi D024: Difference between revisions
From Bioblast
No edit summary |
No edit summary |
||
Line 6: | Line 6: | ||
|SUIT number=D024_1GM;2D;2c;3S;4U;5Rot;6Ama | |SUIT number=D024_1GM;2D;2c;3S;4U;5Rot;6Ama | ||
}} | }} | ||
::: [[MitoPedia: SUIT]]ย ย | ::: '''[[MitoPedia: SUIT]]''' ย | ||
::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS(GM) | ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS(GM) | ||
::: '''[[SUIT protocol pattern]]:''' diametral 1GM;2D;2c;3S;4U;5Rot | ::: '''[[SUIT protocol pattern]]:''' diametral 1GM;2D;2c;3S;4U;5Rot |
Revision as of 13:30, 6 March 2019
Description
Abbreviation: NS(GM)
Reference: A pfi: permeabilized fibers - SUIT-011
SUIT number: D024_1GM;2D;2c;3S;4U;5Rot;6Ama
O2k-Application: O2
- MitoPedia: SUIT
- SUIT-category: NS(GM)
- SUIT protocol pattern: diametral 1GM;2D;2c;3S;4U;5Rot
Communicated by Doerrier C and Gnaiger E (last update 2019-02-11)
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
|
3S | GMSP | NS | CI&II | 1GM;2D;2c;3S
|
4U | GMSE | NS | CI&II | 1GM;2D;2c;3S;4U
|
5Rot | SE | S | CII | 1GM;2D;2c;3S;4U;5Rot
|
6Ama | ROX | 1GM;2D;2c;3S;4U;5Rot;6Ama
|
Step | Respiratory state | Pathway control | ET-Complex | Comment |
---|---|---|---|---|
## AsTm | AsTmE | CIV | CIV | |
## Azd | CHB |
- Bioblast links: SUIT protocols - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- 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
- Comparison of GM- with PM-capacity yields important information on N-pathway respiratory control upstream of CI (Lemieux et al. 2017; Votion et al. 2012).
- A succinate concentration of >10 mM may be required for saturating SE capacity.
- Rox might be inhibited slightly further by inhibition of CIV by cyanide (KCN; 1 ฮผM). But cyanide inhibits not only CIV, but also catalase and other oxygenases involved in ROX.
- + NS-OXPHOS capacity provides a physiologically relevant estimate of maximum mitochondrial respiratory capacity.
- + Glutamate is easier to prepare compared to pyruvate.
- + Application of the cytochrome c test early in the protocol ensures comparability of all states in case of any effect of c.
- + Reasonable duration of the experiment.
- - GM and PM yield typically identical fluxes in human skeletal muscle fibres. However, PM is the superior alternative to GM: the fraction of the N-pathway is lower and of the S-pathway is higher with GM compared to PM (GMP is inhibited by the CII inhibitor malonic acid to a larger extent than PMP). PM, therefore, yields a more sensitive assay for the diagnosis of injuries in the N-pathway, since an impairment of N-pathway capacity can be compensated partially by activation of the S-pathway. This is a disadvantage compared to SUIT-004 and SUIT-008 for diagnosis of N-capacity.
- - To detect an additive effect of P after GMP, pyruvate would have to be added as step 3 (before S). However, inhibition of respiration was observed after titration of P (5 mM) 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
- GM and PM yield typically identical fluxes in human skeletal muscle fibres.
- SUIT-004 1PM;2D;3U;4S;5Rot-
- SUIT-008 1PM;2D;3G;4S;5U;6Rot-
- SUIT-001 1PM;2D;3U;4G;5S;6Oct;7Rot;8Gp-
- 1PGM;2D;3S;4U;5Rot-
References
Year | Reference | Organism | Tissue;cell | |
---|---|---|---|---|
Votion 2012 PLoS One | 2012 | Votion DM, Gnaiger E, Lemieux H, Mouithys-Mickalad A, Serteyn D (2012) Physical fitness and mitochondrial respiratory capacity in horse skeletal muscle. PLoS One 7:e34890. https://doi.org/10.1371/journal.pone.0034890 | Horse | Skeletal muscle |
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry
Labels:
SUIT-011