Difference between revisions of "Kalbacova 2003 Cytometry"
| Line 5: | Line 5: | ||
|year=2003 | |year=2003 | ||
|journal=Cytometry | |journal=Cytometry | ||
|abstract=Background: Determination of mitochondrial membrane | |abstract='''Background:''' Determination of mitochondrial membrane potential ((m) is widely used to characterize cellular metabolism, viability, and apoptosis. Changes of ΔΨm induced by inhibitors of oxidative phosphorylation characterize | ||
potential ((m) is widely used to characterize cellular | respective contributions of mitochondria and glycolysis to adenosine triphosphate (ATP) synthesis. | ||
metabolism, viability, and apoptosis. Changes of ΔΨm | '''Methods:''' ΔΨm in BSC-40 and HeLa G cell lines was determined by flow cytometry and spectrofluorometry. Its changes induced by specific mitochondrial inhibitors were evaluated using 3,3 ΔΨ-dihexyloxacarbocyanine iodide | ||
induced by inhibitors of oxidative phosphorylation characterize | (DiOC6(3)), tetramethylrhodamine ethyl ester, and Mito-Tracker Red. Mitochondrial function was further characterized by oxygen consumption. | ||
respective contributions of mitochondria and glycolysis | '''Results:''' Inhibition of respiration by antimycin A or uncoupling | ||
to adenosine triphosphate (ATP) synthesis. | of mitochondria by FCCP decreased ΔΨm in both cell lines. Inhibition of ATP production by oligomycin or atractyloside induced a moderate decrease of ΔΨm | ||
Methods: ΔΨm in BSC-40 and HeLa G cell lines was | in HeLa G cells and an increase of ΔΨm in BSC-40 cells. Statistically significant differences in ΔΨm between the two cell lines were found with both flow cytometry and spectrofluorometry. Respirometry showed higher basal | ||
determined by flow cytometry and spectrofluorometry. Its | |||
changes induced by specific mitochondrial inhibitors | |||
were evaluated using 3,3 ΔΨ-dihexyloxacarbocyanine iodide | |||
(DiOC6(3)), tetramethylrhodamine ethyl ester, and Mito- | |||
Tracker Red. Mitochondrial function was further characterized | |||
by oxygen consumption. | |||
Results: Inhibition of respiration by antimycin A or uncoupling | |||
of mitochondria by FCCP decreased ΔΨm in | |||
both cell lines. Inhibition of ATP production by oligomycin | |||
or atractyloside induced a moderate decrease of ΔΨm | |||
in HeLa G cells and an increase of ΔΨm in BSC-40 cells. | |||
Statistically significant differences in ΔΨm between the | |||
two cell lines were found with both flow cytometry and | |||
spectrofluorometry. Respirometry showed higher basal | |||
and FCCP-stimulated respiration in BSC-40 cells. | and FCCP-stimulated respiration in BSC-40 cells. | ||
Conclusion: Changes of ΔΨm and oxygen consumption | '''Conclusion:''' Changes of ΔΨm and oxygen consumption | ||
showed that BSC-40 cells are more sensitive than HeLa G | showed that BSC-40 cells are more sensitive than HeLa G | ||
cells to inhibitors of mitochondrial function, suggesting | cells to inhibitors of mitochondrial function, suggesting | ||
| Line 35: | Line 21: | ||
ones by spectrofluorometry. Cytometry Part A 52A: | ones by spectrofluorometry. Cytometry Part A 52A: | ||
110–116, 2003. | 110–116, 2003. | ||
|keywords=monkey | |||
}} | }} | ||
{{Labeling | {{Labeling | ||
|instruments=Oxygraph-2k | |instruments=Oxygraph-2k | ||
|injuries=Cancer; Apoptosis; Cytochrome c | |injuries=Cancer; Apoptosis; Cytochrome c | ||
|organism=Other Mammal | |||
|tissues=Blood Cell; Suspension Culture | |tissues=Blood Cell; Suspension Culture | ||
|preparations=Intact Cell; Cultured; Primary | |||
|kinetics=Inhibitor; Uncoupler | |kinetics=Inhibitor; Uncoupler | ||
|topics=Respiration; OXPHOS; ETS Capacity, Coupling; Membrane Potential, Ion Homeostasis, Substrate; Glucose; TCA Cycle | |topics=Respiration; OXPHOS; ETS Capacity, Coupling; Membrane Potential, Ion Homeostasis, Substrate; Glucose; TCA Cycle | ||
|additional=Spectrophotometry | |additional=Spectrophotometry, Spectrofluorimetry | ||
}} | }} | ||
Revision as of 13:29, 6 September 2011
| Kalbacova M, Vrbacky M, Drahota Z, Melkova Z (2003) Comparison of the effect of mitochondrial inhibitors on mitochondrial membrane potential in two different cell lines using flow cytometry and spectrofluorometry. Cytometry 52A: 110-116. |
Kalbacova M, Vrbacky M, Drahota Z, Melkova Z (2003) Cytometry
Abstract: Background: Determination of mitochondrial membrane potential ((m) is widely used to characterize cellular metabolism, viability, and apoptosis. Changes of ΔΨm induced by inhibitors of oxidative phosphorylation characterize respective contributions of mitochondria and glycolysis to adenosine triphosphate (ATP) synthesis. Methods: ΔΨm in BSC-40 and HeLa G cell lines was determined by flow cytometry and spectrofluorometry. Its changes induced by specific mitochondrial inhibitors were evaluated using 3,3 ΔΨ-dihexyloxacarbocyanine iodide (DiOC6(3)), tetramethylrhodamine ethyl ester, and Mito-Tracker Red. Mitochondrial function was further characterized by oxygen consumption. Results: Inhibition of respiration by antimycin A or uncoupling of mitochondria by FCCP decreased ΔΨm in both cell lines. Inhibition of ATP production by oligomycin or atractyloside induced a moderate decrease of ΔΨm in HeLa G cells and an increase of ΔΨm in BSC-40 cells. Statistically significant differences in ΔΨm between the two cell lines were found with both flow cytometry and spectrofluorometry. Respirometry showed higher basal and FCCP-stimulated respiration in BSC-40 cells. Conclusion: Changes of ΔΨm and oxygen consumption showed that BSC-40 cells are more sensitive than HeLa G cells to inhibitors of mitochondrial function, suggesting that BSC-40 cells are more dependent than HeLa G cells on aerobic ATP production. Determination of ΔΨm changes by flow cytometry exhibited greater sensitivity than the ones by spectrofluorometry. Cytometry Part A 52A: 110–116, 2003. • Keywords: monkey
Labels:
Stress:Cancer; Apoptosis; Cytochrome c"Cancer; Apoptosis; Cytochrome c" is not in the list (Cell death, Cryopreservation, Ischemia-reperfusion, Permeability transition, Oxidative stress;RONS, Temperature, Hypoxia, Mitochondrial disease) of allowed values for the "Stress" property. Organism: Other Mammal"Other Mammal" is not in the list (Human, Pig, Mouse, Rat, Guinea pig, Bovines, Horse, Dog, Rabbit, Cat, ...) of allowed values for the "Mammal and model" property. 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., Ion Homeostasis"Ion Homeostasis" 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
Spectrophotometry, Spectrofluorimetry
