Huetter 2004 Biochem J
| Hütter E, Renner K, Pfister G, Stöckl P, Jansen-Dürr P, Gnaiger E (2004) Senescence-associated changes in respiration and oxidative phosphorylation in primary human fibroblasts. Biochem J 380:919-28. |
» PMID: 15018610 Open Access; pdf
Huetter E, Renner K, Pfister G, Stoeckl P, Jansen-Duerr P, Gnaiger E (2004) Biochem J
Abstract: Limitation of lifespan in replicative senescence is related to oxidative stress, which is probably both the cause and consequence of impaired mitochondrial respiratory function. The respiration of senescent human diploid fibroblasts was analysed by high-resolution respirometry. To rule out cell-cycle effects, proliferating and growth-arrested young fibroblasts were used as controls. Noncoupled respiration, as normalized to citrate synthase activity, remained unchanged, reflecting a constant mitochondrial electron transfer system (ETS) capacity. Oligomycin-inhibited LEAK respiration, however, was significantly increased in mitochondria of senescent cells, indicating a lower coupling of electron transfer to phosphorylation of ADP to ATP. In contrast, growth-arrested young fibroblasts exhibited a higher coupling control compared with proliferating controls. In intact cells, partial uncoupling (dyscoupling) may lead to either decreased oxidative ATP production or a compensatory increase in ROUTINE respiration. To distinguish between these alternatives, we subtracted oligomycin-inhibited respiration from ROUTINE respiration, which allowed us to determine the part of respiratory activity coupled with ATP production. Despite substantial differences in the respiratory acceptor control ratio, ranging from 4 to 11 in the different experimental groups, a fixed proportion of ETS capacity was maintained for coupled oxidative phosphorylation in all the experimental groups. This finding indicates that the senescent cells fully compensate for increased proton leakage by enhanced electron-transfer activity in the ROUTINE state. These results provide a new insight into age-associated defects in mitochondrial function and compensatory mechanisms in intact cells. • Keywords: Aging, Coupling state, Mitochondria, Oxidative stress, Primary human fibroblast, Respiration, Senescence.
• O2k-Network Lab: AT Innsbruck Gnaiger E, AT Innsbruck Jansen-Duerr P
Labels: MiParea: Respiration, mt-Biogenesis;mt-density
Pathology: Aging;senescence
Stress:Oxidative stress;RONS
Organism: Human
Preparation: Intact cells Enzyme: Marker enzyme Regulation: Coupling efficiency;uncoupling, mt-Membrane potential, Uncoupler Coupling state: LEAK, ROUTINE, ETS"ETS" is not in the list (LEAK, ROUTINE, OXPHOS, ET) of allowed values for the "Coupling states" property.
HRR: Oxygraph-2k
Coupling control protocol and nomenclature
- CCP: CCP Huetter 2004 Biochem J
- An update on nomenclature
- State 4o »LEAK state with oligomycin
- State 3u, uncoupled »ETS capacity
- j(R-4o)/3u »NetROUTINE control ratio, ≈R/E = (R-L)/E
- Ro »Rot
- AA »Ama
Comment
- In the context of another study (MiP2005 Abstract), we compared respiration of intact and permeabilized fibroblasts with excellent results. Therefore, permeabilized cells provide a validated model to extend mitochondrial studies of senescence. In fact, the fibroblasts in the present study were permeabilized for the JC1 experiments, to eliminate the confounding effect of the plasma membrane potential.
