Kuznetsov 2002 Anal Biochem: Difference between revisions
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{{Labeling | {{Labeling | ||
|area=Respiration, Instruments;methods, mt-Biogenesis;mt-density, mt-Medicine | |area=Respiration, Instruments;methods, mt-Biogenesis;mt-density, mt-Medicine | ||
|injuries=Ischemia-reperfusion | |||
|organism=Pig | |organism=Pig | ||
|tissues=Liver | |tissues=Liver | ||
|preparations=Permeabilized tissue | |preparations=Permeabilized tissue | ||
|enzymes=Marker enzyme | |enzymes=Marker enzyme | ||
|couplingstates=LEAK, OXPHOS | |couplingstates=LEAK, OXPHOS | ||
|pathways=N, S, CIV, ROX | |pathways=N, S, CIV, ROX | ||
|instruments=Oxygraph-2k | |instruments=Oxygraph-2k | ||
|additional=BEC 2020.2 | |||
|discipline=Mitochondrial Physiology, Biomedicine | |discipline=Mitochondrial Physiology, Biomedicine | ||
}} | }} | ||
==Scientific service== | ==Scientific service== | ||
* [[Tissue storage]] | ::::* [[Tissue storage]] | ||
Revision as of 00:51, 2 January 2021
| Kuznetsov AV, Strobl D, Ruttmann E, Königsrainer A, Margreiter R, Gnaiger E (2002) Evaluation of mitochondrial respiratory function in small biopsies of liver. Anal Biochem 305:186-94. |
Kuznetsov AV, Strobl D, Ruttmann E, Koenigsrainer A, Margreiter R, Gnaiger E (2002) Anal 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 Organism: Pig Tissue;cell: Liver Preparation: Permeabilized tissue Enzyme: Marker enzyme
Coupling state: LEAK, OXPHOS Pathway: N, S, CIV, ROX HRR: Oxygraph-2k
BEC 2020.2
