Difference between revisions of "SUIT-003 O2 ce D028"
From Bioblast
Line 1: | Line 1: | ||
{{MitoPedia | {{MitoPedia | ||
|abbr= | |abbr=S CCP permeability test | ||
|description=[[File:ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama.png| | |description=[[File:ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama.png|500px]] | ||
|info='''B''' '''[[SUIT-003]]''' [[coupling control protocol]] with succinate for permeability test | |info='''B''' '''[[SUIT-003]]''' [[coupling control protocol]] with succinate for permeability test | ||
|SUIT number=D028_ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama | |SUIT number=D028_ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama | ||
|application=O2 | |application=O2 | ||
}} | }} | ||
::: '''[[SUIT protocol pattern]]:''' ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama | |||
::: '''[[SUIT protocol pattern]]:''' | |||
__TOC__ | __TOC__ | ||
Communicated by [[Doerrier C]], [[Iglesias-Gonzalez J]] and [[Gnaiger E]] (last update 2019-06-06) | |||
{{Template: SUIT-003 Ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama}} | {{Template: SUIT-003 Ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama}} | ||
Line 35: | Line 24: | ||
== ceR,L,E format == | == ceR,L,E format == | ||
[[File:CCP(S)04(R,L,E).jpg| | [[File:CCP(S)04(R,L,E).jpg|500px]] In the diagrams of coupling control protocols for intact cells (CCP), the steps may be replaced by the respiratory states. | ||
:::* '''ce4Rot,ce5S;ce6Ama''' | :::* '''ce4Rot,ce5S;ce6Ama''' | ||
Line 41: | Line 30: | ||
:::: '''ce5S''': Succinate is not permeable through the plasma membrane of many intact cells ([[Steinlechner-Maran 1997 Transplantation]]). If succinate is accessible to rotenone-inhibited uncoupled mitochondria in dead cells (dce), respiration will be stimulated. | :::: '''ce5S''': Succinate is not permeable through the plasma membrane of many intact cells ([[Steinlechner-Maran 1997 Transplantation]]). If succinate is accessible to rotenone-inhibited uncoupled mitochondria in dead cells (dce), respiration will be stimulated. | ||
:::: '''ce6Ama''': Inhibition of CIII by antimycin A inhibits succinate-stimulated respiration ([[Stadlmann_2006_Cell Biochem Biophys]]). | :::: '''ce6Ama''': Inhibition of CIII by antimycin A inhibits succinate-stimulated respiration ([[Stadlmann_2006_Cell Biochem Biophys]]). | ||
== References == | |||
{{#ask:[[Category:Publications]] [[Additional label::SUIT-003 Ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama]] | |||
|?Was published in year=Year | |||
|?Has title=Reference | |||
|?Mammal and model=Organism | |||
|?Tissue and cell=Tissue;cell | |||
|format=broadtable | |||
|limit=5000 | |||
|offset=0 | |||
|sort=Was published in year | |||
|order=descending | |||
}} | |||
{{MitoPedia concepts | {{MitoPedia concepts |
Revision as of 13:44, 6 June 2019
Description
Abbreviation: S CCP permeability test
Reference: B SUIT-003 coupling control protocol with succinate for permeability test
SUIT number: D028_ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama
O2k-Application: O2
- SUIT protocol pattern: ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama
Communicated by Doerrier C, Iglesias-Gonzalez J and Gnaiger E (last update 2019-06-06)
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
ce1 | ROUTINE | ce1
| ||
ce2Omy | L | ce1;ce2Omy
| ||
ce3U | E | ce1;ce2Omy;ce3U
| ||
ce4Rot | ROX | ce1;ce2Omy;ce3U;ce4Rot
|
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
ce5S | ce1;ce2Omy;ce3U;ce4Rot;ce5S
| |||
ce6Ama | ROX | ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama
|
- Bioblast links: SUIT protocols - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- Coupling control
- Pathway control
- Main fuel substrates
- » Glutamate, G
- » Glycerophosphate, Gp
- » Malate, M
- » Octanoylcarnitine, Oct
- » Pyruvate, P
- » Succinate, S
- Main fuel substrates
- Glossary
Strengths and limitations
- The SUIT-003 Ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama is a cell coupling control protocol. The total cell count (Nce) is the sum of viable cells (Nvce) and dead cells (Ndce).
- This protocol can be extended with the cell viability test module (See CCV SUIT-003 O2 ce-pce D020).
- For determination of respirometric viability index, it is assumed that both viable and dead cells contain functional mitochondria.
- + With the extended module, S-ET capacity can be assessed after the performance of the coupling control protocol, as well as outer mt-membrane integrity with the addition of cytochrome c.
- + CIV activity can also be measured after the cell permeability test module.
- + Flexibility of experiment duration: from 1 hour (cell coupling control protocol) to 2 hours (permeability test module and CIV module).
- - Cells with a cell membrane succinate transporter are not suitable for the viability or permeability test.
- - The addition of oligomycin has to be carefully tested to avoid inhibitory effects in the evaluation of the ET-capacity.
- - Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler.
ceR,L,E format
In the diagrams of coupling control protocols for intact cells (CCP), the steps may be replaced by the respiratory states.
- ce4Rot,ce5S;ce6Ama
- ce4Rot: In many intact cells ROX measured after titration of rotenone (ce4Rot) is not or only slightly further inhibited by subsequent titration of antimycin A.
- ce5S: Succinate is not permeable through the plasma membrane of many intact cells (Steinlechner-Maran 1997 Transplantation). If succinate is accessible to rotenone-inhibited uncoupled mitochondria in dead cells (dce), respiration will be stimulated.
- ce6Ama: Inhibition of CIII by antimycin A inhibits succinate-stimulated respiration (Stadlmann_2006_Cell Biochem Biophys).
References
MitoPedia concepts: SUIT protocol, SUIT B, Find
MitoPedia methods:
Respirometry