Kuznetsov 2002 Anal Biochem: Difference between revisions
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|title=Kuznetsov AV, Strobl D, Ruttmann E, KΓΆnigsrainer A, Margreiter R, Gnaiger E (2002) Evaluation of mitochondrial respiratory function in small biopsies of liver. Analyt Biochem 305: 186-94. | |title=Kuznetsov AV, Strobl D, Ruttmann E, KΓΆnigsrainer A, Margreiter R, Gnaiger E (2002) Evaluation of mitochondrial respiratory function in small biopsies of liver. Analyt Biochem 305:186-94. | ||
|info=[http://www.ncbi.nlm.nih.gov/pubmed/12054447 PMID: 12054447] | |info=[http://www.ncbi.nlm.nih.gov/pubmed/12054447 PMID: 12054447] | ||
|authors=Kuznetsov AV, Strobl D, Ruttmann E, Koenigsrainer A, Margreiter R, Gnaiger E | |authors=Kuznetsov AV, Strobl D, Ruttmann E, Koenigsrainer A, Margreiter R, Gnaiger E |
Revision as of 23:53, 13 December 2014
Kuznetsov AV, Strobl D, Ruttmann E, KΓΆnigsrainer A, Margreiter R, Gnaiger E (2002) Evaluation of mitochondrial respiratory function in small biopsies of liver. Analyt Biochem 305:186-94. |
Kuznetsov AV, Strobl D, Ruttmann E, Koenigsrainer A, Margreiter R, Gnaiger E (2002) Analyt Biochem
Abstract: Mitochondrial respiratory function was studied in permeabilized pig liver biopsies. The cell membrane was permeabilized mechanically in tissue samples of 2β7 mg, for application of a standardized substrate/inhibitor titration protocol in high-resolution respirometry. Specific respirometric tests demonstrated complete plasma membrane permeabilization and accessibility of substrates to intact mitochondria. High respiratory adenylate control ratios and cytochrome c conservation in the tissue preparation were comparable or even better than in isolated mitochondria. Citrate synthase and cytochrome c oxidase activities remained at 85% of controls after up to 98 h storage of liver tissue at 0Β°C in histidineβtryptophanβketoglutarate solution. Multiple mitochondrial defects, however, were indicated after 48 h cold storage by the decline in respiratory capacity, which was lowered to a larger extent with complex I substrates compared to respiration with substrates for complex II or IV, measured in the absence of cytochrome c. After prolonged ischemia, the adenylate control ratio was significantly reduced, and cytochrome c depletion was detected by the stimulatory effect of cytochrome c. High-resolution respirometry allows the assessment of mitochondrial function in a few milligrams of permeabilized liver tissue, without isolation of mitochondria. This provides a basis for the analysis of mitochondrial function in human liver biopsies. β’ Keywords: Mitochondrial respiration, Pig liver tissue, Cell membrane permeabilization, Cold ischemia, Mitochondrial injury
β’ O2k-Network Lab: AT_Innsbruck_Gnaiger E, AT Innsbruck OROBOROS
Labels: MiParea: Respiration, Instruments;methods, mt-Biogenesis;mt-density, mt-Medicine
Stress:Ischemia-Reperfusion; Preservation"Ischemia-Reperfusion; Preservation" 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: Pig Tissue;cell: Liver Preparation: Permeabilized tissue Enzyme: Marker Enzyme"Marker Enzyme" is not in the list (Adenine nucleotide translocase, Complex I, Complex II;succinate dehydrogenase, Complex III, Complex IV;cytochrome c oxidase, Complex V;ATP synthase, Inner mt-membrane transporter, Marker enzyme, Supercomplex, TCA cycle and matrix dehydrogenases, ...) of allowed values for the "Enzyme" property.
Coupling state: LEAK, OXPHOS
HRR: Oxygraph-2k