MiPNet08.09 CellRespiration: Difference between revisions
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{{Publication | {{Publication | ||
|title=Gnaiger E, Renner K. High-resolution respirometry with cultured cells: A demonstration experiment. Oxygraph-2k workshop report, Schroecken, Austria, March 2003. Mitochondr Physiol Network 08.09. | |title=Gnaiger E, Renner K. High-resolution respirometry with cultured cells: A demonstration experiment. Oxygraph-2k workshop report, Schroecken, Austria, March 2003. Mitochondr Physiol Network 08.09. | ||
|info=[http://www.oroboros.at/ | |info=[http://www.oroboros.at/?protocols_cell_pcp MiPNet08.09] | ||
|year=* | |year=* | ||
|journal=Mitochondr Physiol Network | |journal=Mitochondr Physiol Network | ||
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}} | }} | ||
{{Labeling | {{Labeling | ||
|area=Respiration, Instruments and methods | |||
|organism=Human | |organism=Human | ||
|tissues=Blood cells | |tissues=Blood cells | ||
|preparations=Intact cells | |preparations=Intact cells | ||
|couplingstates=LEAK, ROUTINE, ETS | |couplingstates=LEAK, ROUTINE, ETS | ||
|instruments=Oxygraph-2k, Protocol | |instruments=Oxygraph-2k, Protocol | ||
|additional=O2k-Demo | |additional=O2k-Demo | ||
|discipline=Mitochondrial Physiology | |discipline=Mitochondrial Physiology | ||
|articletype=Protocol; Manual, MiPNet-online Publication | |articletype=Protocol; Manual, MiPNet-online Publication | ||
}} | }} |
Revision as of 11:26, 10 August 2013
Gnaiger E, Renner K. High-resolution respirometry with cultured cells: A demonstration experiment. Oxygraph-2k workshop report, Schroecken, Austria, March 2003. Mitochondr Physiol Network 08.09. |
ยป MiPNet08.09
Abstract: An experiment on cellular respiration is reported from an Oxygraph-2k workshop on high-resolution respirometry. Leukemia cells were incubated at a density of 1 million cells/ml in 2 ml culture medium in two O2k chambers operated in parallel. Cellular ROUTINE respiration, JR, resulted in volume-specific oxygen consumption of 20 pmols-1ml-1. Oxygen concentration changed by merely 6.4 and 6.5 ฮผM in the two O2k chambers over a period of 5 min (<1% air saturation per minute). Inhibition by oligomycin (JL), and rotenone (residual oxygen consumption, JROX; after uncoupling) reduced respiration to 5 and 1 pmols1ml-1, while inducing the non-coupled state by FCCP revealed the capacity of the electron transfer system (ETS) at JE of 50 pmols-1ml-1. The ROUTINE flux control ratio, R/E, was 0.39 (uncoupling control ratio, UCR=E/R=2.6), and the LEAK flux control ratio, L/E, was 0.09 (respiratory control ratio, RCR=E/L=12.0). This indicates tight coupling of OXPHOS, and a large respiratory excess capacity over ROUTINE respiration. The net ROUTINE flux control ratio, netR=(R-L)/E was 0.30, indicating that 30% of ETS capacity was activated for ATP production.
โข O2k-Network Lab: AT_Innsbruck_OROBOROS
Labels: MiParea: Respiration, Instruments and methods"Instruments and methods" is not in the list (Respiration, Instruments;methods, mt-Biogenesis;mt-density, mt-Structure;fission;fusion, mt-Membrane, mtDNA;mt-genetics, nDNA;cell genetics, Genetic knockout;overexpression, Comparative MiP;environmental MiP, Gender, ...) of allowed values for the "MiP area" property.
Organism: Human
Tissue;cell: Blood cells
Preparation: Intact cells
Coupling state: LEAK, ROUTINE, ETS"ETS" is not in the list (LEAK, ROUTINE, OXPHOS, ET) of allowed values for the "Coupling states" property.
HRR: Oxygraph-2k, Protocol"Protocol" is not in the list (Oxygraph-2k, TIP2k, O2k-Fluorometer, pH, NO, TPP, Ca, O2k-Spectrophotometer, O2k-Manual, O2k-Protocol, ...) of allowed values for the "Instrument and method" property.
O2k-Demo