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Difference between revisions of "SUIT-002 O2 ce-pce D007a"

From Bioblast
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{{MitoPedia
{{MitoPedia
|application=O2
|abbr=FNSGp(Oct,PGM)
|abbr=FNSGp(Oct,PGM)
|description=[[File:1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot-.png|400px]]
|description=[[File:1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot-.png|400px]]
|SUIT number=D07a_ce1;1Dig;1D;2M;3Oct;3c;4M;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd  
|SUIT number=D07a_ce1;1Dig;1D;2M;3Oct;3c;4M;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd  
|info='''A: SUIT RP2 specific for PBMC and platelets cells''' - [[SUIT-002]]
|info='''A: SUIT RP2 specific for PBMC and platelets cells''' - [[SUIT-002]]
}}
|application=O2
{{MitoPedia concepts
|mitopedia concept=SUIT protocol
}}
}}
::: [[MitoPedia: SUIT]] - '''[[SUIT reference protocol]] RP2'''
::: [[MitoPedia: SUIT]] - '''[[SUIT reference protocol]] RP2'''
::: '''[[Categories of SUIT protocols|SUIT-category]]:''' FNSGp(Oct,PGM)
::: '''[[Categories of SUIT protocols|SUIT-category]]:''' FNSGp(Oct,PGM)
::: '''[[SUIT protocol pattern]]:''' diametral
::: '''[[SUIT protocol pattern]]:''' diametral 1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot


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{{Template:SUIT-002}}
{{Template:SUIT-002}}
{{MitoPedia concepts
|mitopedia concept=SUIT protocol
}}
{{MitoPedia methods
|mitopedia method=Respirometry
}}

Revision as of 09:28, 15 January 2019


high-resolution terminology - matching measurements at high-resolution


SUIT-002 O2 ce-pce D007a

Description

1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot-.png

Abbreviation: FNSGp(Oct,PGM)

Reference: A: SUIT RP2 specific for PBMC and platelets cells - SUIT-002

SUIT number: D07a_ce1;1Dig;1D;2M;3Oct;3c;4M;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd

O2k-Application: O2

MitoPedia: SUIT - SUIT reference protocol RP2
SUIT-category: FNSGp(Oct,PGM)
SUIT protocol pattern: diametral 1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot


Harmonized SUIT protocols

RP1 and RP2

SUIT RP1 SUIT-RP1 SUIT RP2 SUIT-RP2

SUIT RP1: 1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp;9Ama;10AsTm;11Azd
SUIT RP2: 1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd

RP1&RP2.png Harmonization between RP1 and RP2

RP1 and RP2 in cells

RP2 can be assessed in cells after the selective plasma membrane permeabilization. Digitonin is a mild detergent that permeabilizes plasma membranes. The optimum effective digitonin concentration for complete plasma membrane permeabilization of cultured cells can be determined directly in a respirometric protocol (see Digitonin). The use of intact cells allows the possibility to evaluate respiration in two different modules: intact cells and permeabilized cells. ROUTINE respiration (ce1) is obtained before digitonin is employed, whereas after the use of digitonin we can address differentet Electron transfer-pathway statea.

SUIT RP1 in cells: ce1;1Dig;1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp;9Ama;10Tm;11Azd
SUIT RP2 in cells: ce1;1Dig;1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azdd

Reference protocol RP1&RP2 pce.png Harmonization between RP1 and RP2 in cells


RP1 and RP2 in PBMCs and PTLs

In PBMCs and PLTs the reference protocol RP1 is performed in the absense of Oct: SUIT 1 for PBMCs and PTLs

SUIT RP1 in PBMCs and PTLsSUIT RP1 for PBMCs and PLTs:1PM;2D;2c;3U;4G;5S;6Rot;7Gp;8Ama;9AsTm;10Azd
SUIT RP2 PBMCs and PLTs: 1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd

Reference protocol RP1&RP2 pce PBMCs PTLs.png Harmonization between RP1 and RP2 in PBMC and PTL cells


References

MitoPedia: SUIT

Steps and respiratory states

1D;2M.1;3Oct;4M2;5P;6G;7S;8Gp;9U;10Rot-.png

Step State Pathway Q-junction Comment - Events (E) and Marks (M)
1D REN 1D
  • ADP is added to stimulate the consumption of endogenous fuel-substrates.


2M.1 1D;2M.1
3Oct OctMP F FAO 1D;2M.1;3Oct
  • Respiratory stimulation of the FAO-pathway, F, by fatty acid, FA, in the presence of malate, M. Malate is a type N substrate (N), required for the F-pathway. The FA concentration has to be optimized to saturate the F-pathway, without inhibiting or uncoupling respiration.
3c OctMcP F FAO 1D;2M.1;3Oct;3c
  • Respiratory stimulation of the FAO-pathway, F, by fatty acid, FA, in the presence of malate, M. Malate is a type N substrate (N), required for the F-pathway. The FA concentration has to be optimized to saturate the F-pathway, without inhibiting or uncoupling respiration.
  • OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
  • Addition of cytochrome c yields a test for integrity of the mtOM (cytochrome c control efficiency). Stimulation by added cytochrome c would indicate an injury of the mtOM and limitation of respiration in the preceding state without added c due to loss of cytochrome c. Typically, cytochrome c is added immediately after the earliest ADP-activation step (OXPHOS capacity P with saturating [ADP]).
4M2 OctMP F(N) FAO 1D;2M.1;3Oct;3c;4M2
5P OctPMP FN FAO&CI 1D;2M.1;3Oct;4M2;5P
  • Respiratory stimulation by simultaneous action of the F-pathway and N-pathway with convergent electron flow in the FN-pathway for evaluation of an additive or inhibitory effect of F.
  • OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
6G OctPGMP FN FAO&CI 1D;2M.1;3Oct;4M2;5P;6G
  • Respiratory stimulation by simultaneous action of the F-pathway and N-pathway with convergent electron flow in the FN-pathway for evaluation of an additive or inhibitory effect of F.
  • OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
7S OctPGMSP FNS FAO&CI&II 1D;2M.1;3Oct;4M2;5P;6G;7S
  • Respiratory stimulation by simultaneous action of the F-pathway, N-pathway, and S-pathway, with convergent electron flow in the FNS-pathway for reconstitution of TCA cycle function and additive or inhibitory effect of F.
  • OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
8Gp OctPGMSGpP FNSGp FAO&CI&II&GpDH 1D;2M.1;3Oct;4M2;5P;6G;7S;8Gp
9U OctPGMSGpE FNSGp FAO&CI&II&GpDH 1D;2M.1;3Oct;4M2;5P;6G;7S;8Gp;9U
10Rot SGpE SGp CII&GpDH 1D;2M.1;3Oct;4M2;5P;6G;7S;8Gp;9U;10Rot
  • Respiratory stimulation by action of succinate and glycerophosphate, Gp, with convergent electron flow in the SGp-pathway (CII&GpDH-linked pathway to the Q-junction).
  • Noncoupled electron transfer state, ET state, with ET capacity E.
11Ama ROX 1D;2M.1;3Oct;4M2;5P;6G;7S;8Gp;9U;10Rot;11Ama
  • Rox is the residual oxygen consumption in the ROX state, due to oxidative side reactions, estimated after addition of antimycin A (inhibitor of CIII). Rox is subtracted from oxygen flux as a baseline for all respiratory states, to obtain mitochondrial respiration (mt).


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MitoPedia concepts: SUIT protocol 


MitoPedia methods: Respirometry