MiPNet08.09 CellRespiration: Difference between revisions
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|authors=Gnaiger E, Renner K | |authors=Gnaiger E, Renner K | ||
|year=2003 | |year=2003 | ||
|journal=Mitochondr. Physiol. Network | |||
|mipnetlab=AT_Innsbruck_OROBOROS | |mipnetlab=AT_Innsbruck_OROBOROS | ||
|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 pmol๏s-1๏ml-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 pmol๏s1๏ml-1, while inducing the non-coupled state by FCCP revealed the capacity of the electron transfer system (ETS) at JE of 50 pmol๏s-1๏ml-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. | |||
|info=[http://www.oroboros.at/index.php?id=protocols_cell_respiration MiPNet08.09] | |info=[http://www.oroboros.at/index.php?id=protocols_cell_respiration MiPNet08.09] | ||
}} | }} | ||
{{Labeling | {{Labeling | ||
|discipline=Mitochondrial Physiology | |discipline=Mitochondrial Physiology | ||
|organism=Human | |organism=Human | ||
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|preparations=Intact Cell; Cultured; Primary | |preparations=Intact Cell; Cultured; Primary | ||
|topics=Respiration; OXPHOS; ETS Capacity, Coupling; Membrane Potential | |topics=Respiration; OXPHOS; ETS Capacity, Coupling; Membrane Potential | ||
|instruments=Oxygraph-2k, DatLab Software; Separate Application, Method, Theory | |||
|articletype=Protocol; Manual, MiPNet-online Publication | |||
}} | }} | ||
[[Category:OroboroPedia]] | [[Category:OroboroPedia]] | ||
Revision as of 11:11, 20 October 2010
| 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
Gnaiger E, Renner K (2003) Mitochondr. Physiol. Network
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:
Organism: Human
Tissue;cell: Blood Cell; Suspension Culture"Blood Cell; Suspension Culture" is not in the list (Heart, Skeletal muscle, Nervous system, Liver, Kidney, Lung;gill, Islet cell;pancreas;thymus, Endothelial;epithelial;mesothelial cell, Blood cells, Fat, ...) of allowed values for the "Tissue and cell" property.
Preparation: Intact Cell; Cultured; Primary"Intact Cell; Cultured; Primary" is not in the list (Intact organism, Intact organ, Permeabilized cells, Permeabilized tissue, Homogenate, Isolated mitochondria, SMP, Chloroplasts, Enzyme, Oxidase;biochemical oxidation, ...) of allowed values for the "Preparation" property.
Regulation: Respiration; OXPHOS; ETS Capacity"Respiration; OXPHOS; ETS Capacity" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property., Coupling; Membrane Potential"Coupling; Membrane Potential" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property.
HRR: Oxygraph-2k, DatLab Software; Separate Application"DatLab Software; Separate Application" 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., Method"Method" 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., Theory
