Difference between revisions of "Gnaiger 1998 BTK-HRR"

Revision as of 20:24, 13 September 2010

Gnaiger E, Kuznetsov AV, Lassnig B, Fuchs A, Reck M, Renner K, Stadlmann S, Rieger G, Margreiter R (1998) High-resolution respirometry. Optimum permeabilization of the cell membrane by digitonin. In BioThermoKinetics in the Post Genomic Era (Larsson C, Påhlman I-L, Gustafsson L, eds) Chalmers Reproservice, Göteborg: 89-95.

» BTK 1998: 89-95

Gnaiger E, Kuznetsov AV, Lassnig B, Fuchs A, Reck M, Renner K, Stadlmann S, Rieger G, Margreiter R (1998)

Abstract: Permeabilization of the cell membrane is becoming an established alternative to the isolation of mitochondria in bioenergetic studies of cultured cells and biopsy samples of muscle. Depending on cell type, 5 to 50 million cells are required for a respirometric measurement of endogenous and permeabilized-cell respiration [1,2], using conventional oxygen monitoring instruments. An order of magnitude less cells are required when using highresolution respirometry [3] (Tab. 1), which provides an up-to-date standard for general and clinical bioenergetics, including the diagnosis of mitochondrial defects, testing of drugs, oxidative stress and hypoxia-reoxygenation injury [4-6]. We provide an overview of (i) the specific features of high-resolution respirometry, (ii) test experiments for instrumental evaluation, and (iii) digitonin titrations for determining optimum concentrations for cell membrane permeabilization in cultured cells.

• O2k-Network Lab: AT_Innsbruck_GnaigerE

Labels:

Organism: Human Tissue;cell: Endothelial; Epithelial; Mesothelial Cell"Endothelial; Epithelial; Mesothelial Cell" 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., Permeabilized Cell or Tissue; Homogenate"Permeabilized Cell or Tissue; Homogenate" 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., Substrate; Glucose; TCA Cycle"Substrate; Glucose; TCA Cycle" 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, TIP2k, 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.

@@ Line 4: / Line 4: @@
 |year=1998
 |mipnetlab=AT_Innsbruck_GnaigerE
-|abstract=Permeabilization of the cell membrane is becoming an established alternative to the isolation of mitochondria in bioenergetic studies of cultured cells and biopsy samples of muscle. Depending on cell type, 5 to 50 million cells are required for a respirometric measurement of endogenous and permeabilized-cell respiration [1,2], using conventional oxygen monitoring instruments. An order of magnitude less cells are required when using highresolution respirometry [3] (Tab. 1), which provides an up-to-date standard for general and clinical bioenergetics, including the diagnosis of mitochondrial defects, testing of drugs, oxidative stress and hypoxia-reoxygenation injury [4-6]. We provide an overview of (i) the specific features of high-resolution respirometry, (ii)
+|abstract=Permeabilization of the cell membrane is becoming an established alternative to the isolation of mitochondria in bioenergetic studies of cultured cells and biopsy samples of muscle. Depending on cell type, 5 to 50 million cells are required for a respirometric measurement of endogenous and permeabilized-cell respiration [1,2], using conventional oxygen monitoring instruments. An order of magnitude less cells are required when using highresolution respirometry [3] (Tab. 1), which provides an up-to-date standard for general and clinical bioenergetics, including the diagnosis of mitochondrial defects, testing of drugs, oxidative stress and hypoxia-reoxygenation injury [4-6]. We provide an overview of (i) the specific features of high-resolution respirometry, (ii) test experiments for instrumental evaluation, and (iii) digitonin titrations for determining optimum concentrations for cell membrane permeabilization in cultured cells.
-test experiments for instrumental evaluation, and (iii) digitonin titrations for determining optimum concentrations for cell membrane permeabilization in cultured cells.
 |info=[http://www.oroboros.at/fileadmin/user_upload/Reprints/1998_Gnaiger_et_al_BTK_HRrespirometry.pdf BTK 1998: 89-95]
 }}