Senescence-associated changes in respiration and oxidative phosphorylation in primary human fibroblasts.
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. Uncoupled respiration, as normalized to citrate synthase activity, remained unchanged, reflecting a constant capacity of the respiratory chain. Oligomycin-inhibited respiration, however, was significantly increased in mitochondria of senescent cells, indicating a lower coupling of electron transport with phosphorylation. In contrast, growth-arrested young fibroblasts exhibited a higher coupling state compared with proliferating controls. In intact cells, partial uncoupling 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 control ratio, ranging from 4 to 11 in the different experimental groups, a fixed proportion of respiratory 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-transport activity in the routine state. These results provide a new insight into age-associated defects in mitochondrial function and compensatory mechanisms in intact cells.
Hutter
Eveline
E
Abteilung Molekular-und Zellbiologie, Institut für Biomedizinische Alternsforschung der Osterreichischen Akademie der Wissenschaften, Rennweg 10, A-6020 Innsbruck, Austria.
Renner
Kathrin
K
Pfister
Gerald
G
Stöckl
Petra
P
Jansen-Dürr
Pidder
P
Gnaiger
Erich
E
D000375:Aging / Q000378:metabolism* / Q000502:physiology; D002452:Cell Count; D002460:Cell Line; D019069:Cell Respiration / Q000502:physiology*; D002950:Citrate (si)-Synthase / Q000378:metabolism; D005347:Fibroblasts / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008564:Membrane Potentials / Q000502:physiology; D008928:Mitochondria / Q000201:enzymology / Q000378:metabolism / Q000502:physiology; D010085:Oxidative Phosphorylation
D007770:L-Lactate Dehydrogenase; D002950:Citrate (si)-Synthase