Difference between revisions of "Kuznetsov 2002 Anal Biochem"
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|preparations=Permeabilized tissue | |preparations=Permeabilized tissue | ||
|enzymes=Marker | |enzymes=Marker enzyme | ||
|injuries=Ischemia- | |injuries=Ischemia-reperfusion;preservation | ||
|couplingstates=LEAK, OXPHOS | |couplingstates=LEAK, OXPHOS | ||
|substratestates=CI, CII, CIV, ROX | |substratestates=CI, CII, CIV, ROX | ||
Revision as of 16:53, 17 February 2015
| Kuznetsov AV, Strobl D, Ruttmann E, Königsrainer 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 2–7 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 0°C in histidine–tryptophan–ketoglutarate 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. • Keywords: Mitochondrial respiration, Pig liver tissue, Cell membrane permeabilization, Cold ischemia, Mitochondrial injury
• 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
Coupling state: LEAK, OXPHOS
HRR: Oxygraph-2k
