Kuznetsov 2002 Anal Biochem: Difference between revisions
(Created page with "{{Publication |title=Kuznetsov AV, Strobl D, Ruttmann E, Königsrainer A, Margreiter R, Gnaiger E (2002) Evaluation of mitochondrial respiratory function in small biopsies of liv...") |
No edit summary |
||
(36 intermediate revisions by 10 users not shown) | |||
Line 1: | Line 1: | ||
{{Publication | {{Publication | ||
|title=Kuznetsov AV, Strobl D, Ruttmann E, Königsrainer A, Margreiter R, Gnaiger E (2002) Evaluation of mitochondrial respiratory function in small biopsies of liver. | |title=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. | ||
|authors=Kuznetsov AV, Strobl D, Ruttmann E, | |info=[http://www.ncbi.nlm.nih.gov/pubmed/12054447 PMID: 12054447] | ||
|authors=Kuznetsov AV, Strobl D, Ruttmann E, Koenigsrainer A, Margreiter R, Gnaiger Erich | |||
|year=2002 | |year=2002 | ||
|journal= | |journal=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. | |||
|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 | |keywords=Mitochondrial respiration, Pig liver tissue, Cell membrane permeabilization, Cold ischemia, Mitochondrial injury | ||
| | |mipnetlab=AT Innsbruck Gnaiger E, AT Innsbruck Oroboros | ||
}} | }} | ||
::::* [[Tissue storage]] | |||
== Cited by == | |||
::* 41 articles in PubMed (2021-12-27) https://pubmed.ncbi.nlm.nih.gov/12054447/ | |||
{{Template:Cited by Gnaiger 2020 BEC MitoPathways}} | |||
{{Labeling | {{Labeling | ||
| | |area=Respiration, Instruments;methods, mt-Biogenesis;mt-density, mt-Medicine | ||
| | |injuries=Ischemia-reperfusion | ||
|organism=Pig | |organism=Pig | ||
|tissues= | |tissues=Liver | ||
|preparations=Permeabilized | |preparations=Permeabilized tissue | ||
| | |enzymes=Marker enzyme | ||
| | |couplingstates=LEAK, OXPHOS | ||
|pathways=N, S, CIV, ROX | |||
|instruments=Oxygraph-2k | |||
|additional=BEC 2020.2 | |||
}} | }} |
Latest revision as of 14:05, 27 December 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 Erich (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
Cited by
- 41 articles in PubMed (2021-12-27) https://pubmed.ncbi.nlm.nih.gov/12054447/
- Gnaiger E (2020) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 5th ed. Bioenerg Commun 2020.2. https://doi.org/10.26124/bec:2020-0002
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