On isolated mitochondria, numerous studies of the relationships between fluxes and their associated forces have led to the description of some properties of the oxidative phosphorylation pathway. However whether such an approach can be applied to understanding the actual situation in intact living cells needs further consideration. In this study on isolated hepatocytes, we describe the dependence of the respiratory rate on the three thermodynamic forces linked to oxidative phosphorylation (i.e. the redox span over the respiratory chain, the electrical potential difference across the inner mitochondrial membrane and the free energy of ATP synthesis reaction). Even if this description is phenomenological and some objections may be raised regarding the relevance of such a bulk-phase force estimation, we present some results showing that the study of flux-force relationships in intact cells may be a helpful approach for understanding the mechanisms by which oxidative phosphorylation activity is changed.

In isolated hepatocytes incubated in hypoosmotic media, a large increase in the mitochondrial volume is not directly involved in the activation of respiration. Moreover, results of the quantification of the various bioenergetic parameters are not in accordance with an activation of the respiratory chain as previously proposed (Halestrap, A.P. (1989) Biochim. Biophys. Acta, 973, 355-382), but point more to an inhibition of respiration. The same respiration rate is obtained in hypoosmolar incubation media in vitro and in situ for a higher overall thermodynamic driving force over the electron transport chain.