Difference between revisions of "Chowdhury 2000 Clin Chim Acta"
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{{Publication | {{Publication | ||
|title=Chowdhury SK, Drahota Z, Floryk D, Calda P, Houstek J | |title=Chowdhury SK, Drahota Z, Floryk D, Calda P, Houstek J (2000) Activities of mitochondrial oxidative phosphorylation enzymes in cultured amniocytes. Clin Chim Acta 298:157-73. | ||
|info=[http://www.ncbi.nlm.nih.gov/pubmed/10876012 PMID: 10876012] | |||
|authors=Chowdhury SK, Drahota Z, Floryk D, Calda P, Houstek J | |authors=Chowdhury SK, Drahota Z, Floryk D, Calda P, Houstek J | ||
|year=2000 | |year=2000 | ||
|journal= | |journal=Clin Chim Acta | ||
|abstract=Amniocytes represent a population of foetal cells that can be used for prenatal diagnosis in families with suspected mitochondrial oxidative phosphorylation (OXPHOS) defects. In this paper, we present a complex protocol for evaluation of the function of mitochondrial OXPHOS enzymes in cultured amniocytes using three independent and complementary methods: (a) spectrophotometry as a tool for determination of the capacities of mitochondrial respiratory-chain enzymes (NADH ubiquinone oxidoreductase, succinate- and glycerophosphate cytochrome c reductase, cytochrome c oxidase and citrate synthase); (b) polarography as a tool for the evaluation of mitochondrial OXPHOS enzyme functions in situ using digitonin-permeabilised amniocytes (rotenone-sensitive oxidation of pyruvate+malate, antimycin A-sensitive oxidation of succinate, KCN-sensitive oxidation of cytochrome c, ADP-activated substrate oxidation) and (c) cytofluorometric determination of tetramethyl rhodamine methyl ester (TMRM) fluorescence in digitonin-permeabilised amniocytes as a sensitive way to determine the mitochondrial membrane potential under steady-state conditions (state 4 with succinate). These protocols are presented together with reference control values using 9–22 independent cultures of amniocytes. | |abstract=Amniocytes represent a population of foetal cells that can be used for prenatal diagnosis in families with suspected mitochondrial oxidative phosphorylation (OXPHOS) defects. In this paper, we present a complex protocol for evaluation of the function of mitochondrial OXPHOS enzymes in cultured amniocytes using three independent and complementary methods: (a) spectrophotometry as a tool for determination of the capacities of mitochondrial respiratory-chain enzymes (NADH ubiquinone oxidoreductase, succinate- and glycerophosphate cytochrome c reductase, cytochrome c oxidase and citrate synthase); (b) polarography as a tool for the evaluation of mitochondrial OXPHOS enzyme functions in situ using digitonin-permeabilised amniocytes (rotenone-sensitive oxidation of pyruvate+malate, antimycin A-sensitive oxidation of succinate, KCN-sensitive oxidation of cytochrome c, ADP-activated substrate oxidation) and (c) cytofluorometric determination of tetramethyl rhodamine methyl ester (TMRM) fluorescence in digitonin-permeabilised amniocytes as a sensitive way to determine the mitochondrial membrane potential under steady-state conditions (state 4 with succinate). These protocols are presented together with reference control values using 9–22 independent cultures of amniocytes. | ||
|keywords=Prenatal diagnosis, Amniocytes, Oxidative phosphorylation, Respiratory-chain enzymes, Mitochondrial membrane potential, TMRM cytofluorometry | |keywords=Prenatal diagnosis, Amniocytes, Oxidative phosphorylation, Respiratory-chain enzymes, Mitochondrial membrane potential, TMRM cytofluorometry | ||
| | |mipnetlab=CZ Prague Houstek J, CA Winnipeg Fernyhough P, CZ Hradec Kralove Cervinkova Z | ||
}} | }} | ||
{{Labeling | {{Labeling | ||
| | |area=Respiration | ||
|tissues=Endothelial; | |organism=Human | ||
| | |tissues=Endothelial;epithelial;mesothelial cell, Fibroblast | ||
|preparations=Permeabilized cells | |||
|couplingstates=LEAK, ROUTINE, OXPHOS | |||
|pathways=N, S, CIV | |||
|instruments=Oxygraph-2k | |instruments=Oxygraph-2k | ||
}} | }} | ||
Latest revision as of 19:08, 1 April 2018
| Chowdhury SK, Drahota Z, Floryk D, Calda P, Houstek J (2000) Activities of mitochondrial oxidative phosphorylation enzymes in cultured amniocytes. Clin Chim Acta 298:157-73. |
Chowdhury SK, Drahota Z, Floryk D, Calda P, Houstek J (2000) Clin Chim Acta
Abstract: Amniocytes represent a population of foetal cells that can be used for prenatal diagnosis in families with suspected mitochondrial oxidative phosphorylation (OXPHOS) defects. In this paper, we present a complex protocol for evaluation of the function of mitochondrial OXPHOS enzymes in cultured amniocytes using three independent and complementary methods: (a) spectrophotometry as a tool for determination of the capacities of mitochondrial respiratory-chain enzymes (NADH ubiquinone oxidoreductase, succinate- and glycerophosphate cytochrome c reductase, cytochrome c oxidase and citrate synthase); (b) polarography as a tool for the evaluation of mitochondrial OXPHOS enzyme functions in situ using digitonin-permeabilised amniocytes (rotenone-sensitive oxidation of pyruvate+malate, antimycin A-sensitive oxidation of succinate, KCN-sensitive oxidation of cytochrome c, ADP-activated substrate oxidation) and (c) cytofluorometric determination of tetramethyl rhodamine methyl ester (TMRM) fluorescence in digitonin-permeabilised amniocytes as a sensitive way to determine the mitochondrial membrane potential under steady-state conditions (state 4 with succinate). These protocols are presented together with reference control values using 9–22 independent cultures of amniocytes. • Keywords: Prenatal diagnosis, Amniocytes, Oxidative phosphorylation, Respiratory-chain enzymes, Mitochondrial membrane potential, TMRM cytofluorometry
• O2k-Network Lab: CZ Prague Houstek J, CA Winnipeg Fernyhough P, CZ Hradec Kralove Cervinkova Z
Labels: MiParea: Respiration
Organism: Human
Tissue;cell: Endothelial;epithelial;mesothelial cell, Fibroblast
Preparation: Permeabilized cells
Coupling state: LEAK, ROUTINE, OXPHOS
Pathway: N, S, CIV
HRR: Oxygraph-2k
