Wagenaars 2014 Abstract IOC 2014-04 Schroecken: Difference between revisions
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{{ | {{Abstract | ||
|title= | |title=Wagenaars J, Willems PHGM, Koopman WJ (2014) Oxygen consumption in OXPHOS-deficient cells. Mitochondr Physiol Network 19.02. | ||
|authors=Wagenaars J, Willems PHGM, Koopman WJ | |||
|authors= | |year=2014 | ||
|year= | |event=[[MiPNet19.02 IOC88]] | ||
|event= | |abstract=Mitochondria are crucially involved in cellular Ca2+ and redox homeostasis and apoptosis induction. Maintenance of mitochondrial function and integrity requires an inside-negative potential difference across the mitochondrial inner membrane. This potential is sustained by four complexes (CI-CIV) of the electron-transport chain (ETC). Together with the ATP-generating FoF1-ATPase (complex V or CV) the ETC complexes constitute the oxidative phosphorylation (OXPHOS) system. CI or NADH:ubiquinone oxidoreductase is the first and largest protein complex of the ETC and couples the oxidation of NADH to the reduction of ubiquinone. | ||
| | In my current research I focus on the analysis of primary skin fibroblasts from patients with mitochondrial OXPHOS disorders (including CI deficiency) and myofibers/myoblasts/myotubes from wildtype (WT) and KO animals with CI deficiency (NDUFS4-/- mice). We have observed that intact fibroblasts from patients with a mild CI deficiency display normal routine O2 consumption, whereas patient fibroblasts with a very severe CI deficiency display reduced routine O2 consumption (threshold effect). Furthermore we observed no differences in maximal (FCCP-induced) respiration in fibroblasts from patients with a mild isolated CI deficiency. To stimulate mitochondrial ATP-linked respiration, cells were treated with the hormone bradykinin during O2 consumption measurements. In this type of experiment the magnitude of the bradykinin-stimulated O2 consumption was less in fibroblasts from patients with isolated CI deficiency than in cells from a healthy individual. | ||
In this workshop I aim to increase my general knowledge of the Oroboros system and its (im)possibilities. I am particularly interested in strategies that can be applied in intact cells to investigate OXPHOS deficiencies. Moreover, the meaning and relevance of “reserve capacity” in the context of intact cells I find particularly interesting. | |||
Cell types: | |||
|mipnetlab= | - Primary patient and animal cells | ||
- Fibroblasts, myoblasts, myofibers | |||
- Intact vs. permeabilized vs. isolated mitochondria | |||
|mipnetlab=NL Nijmegen Koopman WJ | |||
}} | }} | ||
{{ | {{Labeling | ||
|area= | |area=Respiration | ||
| | |injuries=Mitochondrial disease | ||
| | |organism=Human, Mouse | ||
|tissues= | |tissues=Skeletal muscle, Fibroblast | ||
|preparations=Intact cells, Permeabilized cells, Isolated mitochondria | |||
|preparations= | |instruments=Oxygraph-2k | ||
|instruments= | |||
}} | }} | ||
Latest revision as of 15:46, 9 November 2016
| Wagenaars J, Willems PHGM, Koopman WJ (2014) Oxygen consumption in OXPHOS-deficient cells. Mitochondr Physiol Network 19.02. |
Link:
Wagenaars J, Willems PHGM, Koopman WJ (2014)
Event: MiPNet19.02 IOC88
Mitochondria are crucially involved in cellular Ca2+ and redox homeostasis and apoptosis induction. Maintenance of mitochondrial function and integrity requires an inside-negative potential difference across the mitochondrial inner membrane. This potential is sustained by four complexes (CI-CIV) of the electron-transport chain (ETC). Together with the ATP-generating FoF1-ATPase (complex V or CV) the ETC complexes constitute the oxidative phosphorylation (OXPHOS) system. CI or NADH:ubiquinone oxidoreductase is the first and largest protein complex of the ETC and couples the oxidation of NADH to the reduction of ubiquinone. In my current research I focus on the analysis of primary skin fibroblasts from patients with mitochondrial OXPHOS disorders (including CI deficiency) and myofibers/myoblasts/myotubes from wildtype (WT) and KO animals with CI deficiency (NDUFS4-/- mice). We have observed that intact fibroblasts from patients with a mild CI deficiency display normal routine O2 consumption, whereas patient fibroblasts with a very severe CI deficiency display reduced routine O2 consumption (threshold effect). Furthermore we observed no differences in maximal (FCCP-induced) respiration in fibroblasts from patients with a mild isolated CI deficiency. To stimulate mitochondrial ATP-linked respiration, cells were treated with the hormone bradykinin during O2 consumption measurements. In this type of experiment the magnitude of the bradykinin-stimulated O2 consumption was less in fibroblasts from patients with isolated CI deficiency than in cells from a healthy individual. In this workshop I aim to increase my general knowledge of the Oroboros system and its (im)possibilities. I am particularly interested in strategies that can be applied in intact cells to investigate OXPHOS deficiencies. Moreover, the meaning and relevance of “reserve capacity” in the context of intact cells I find particularly interesting. Cell types: - Primary patient and animal cells - Fibroblasts, myoblasts, myofibers - Intact vs. permeabilized vs. isolated mitochondria
• O2k-Network Lab: NL Nijmegen Koopman WJ
Labels: MiParea: Respiration
Stress:Mitochondrial disease Organism: Human, Mouse Tissue;cell: Skeletal muscle, Fibroblast Preparation: Intact cells, Permeabilized cells, Isolated mitochondria
HRR: Oxygraph-2k
