Maintained mitochondrial integrity without oxygen in the anoxia-tolerant crucian carp

Very few vertebrates survive without oxygen (anoxia) for more than a few minutes. Crucian carp (Carassius carassius) are one example, surviving months of anoxia at low temperatures, and we hypothesised that they maintain mitochondrial membrane potential and function. Isolated crucian carp cardiomyocytes indeed maintained mitochondrial membrane potential after blocking complex IV of the electron transport system with cyanide, while those of anoxia-intolerant trout depolarised. When complexes I-III were inhibited, crucian carp mitochondria depolarised, indicating that these complexes need to function during anoxia. Mitochondrial membrane potential depended on reversal of ATP synthase in chemical anoxia, as blocking with cyanide combined with oligomycin to inhibit ATP synthase led to depolarisation. ATP synthase activity was reduced in the heart after 1 week of anoxia in crucian carp, together with a downregulation of ATP synthase subunit gene expression. However, the morphology of cardiac mitochondria was not affected by 1 week of anoxia, even with a large increase in mitofusin 2 mRNA expression. Cardiac citrate synthase activity was not affected by anoxia, while cytochrome c oxidase activity was increased. We show how mitochondria respond to anoxia. A mechanistic understanding of how mitochondrial function can be maintained in anoxia may provide new perspectives to reduce mitochondrial damage in anoxia-sensitive organisms.