Difference between revisions of "SUIT-003 O2 ce D028"
From Bioblast
Line 9: | Line 9: | ||
::: '''[[SUIT protocol pattern]]:''' linear cell [[coupling control protocol]] ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama | ::: '''[[SUIT protocol pattern]]:''' linear cell [[coupling control protocol]] ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama | ||
__TOC__ | __TOC__ | ||
== 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 | |||
}} | |||
{{Template: SUIT-003 Ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama}} | {{Template: SUIT-003 Ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama}} | ||
== Strengths and limitations == | |||
:::* The SUIT-003 Ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama is a cell coupling control protocol. The total cell count (''N''<sub>ce</sub>) is the sum of viable cells (''N''<sub>vce</sub>) and dead cells (''N''<sub>dce</sub>). | |||
:::* 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 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 == | == ceR,L,E format == |
Revision as of 15:55, 21 January 2019
Description
Abbreviation: ceS
Reference: A SUIT-003 coupling control protocol with succinate for permeability test
SUIT number: D###ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama
O2k-Application: O2
- SUIT-category: ceS
- SUIT protocol pattern: linear cell coupling control protocol ce1;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama
References
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 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).
MitoPedia concepts:
SUIT protocol,
SUIT A,
Find
MitoPedia methods:
Respirometry