The increased proton permeability induced by the addition of a synthetic proton translocator to non‐respiring hamster brown‐fat mitochondria is unaffected by purine nucleotide addition. In contrast the permeability induced by fatty acids is inhibited by nucleotide, indicating that fatty acids act at the 32000‐M_r uncoupling protein. Fatty acids lower the affinity of nucleotide binding to the 32000‐M_r protein, but not sufficiently to explain their uncoupling action. The sensitivity of the fatty acid modulation of permeability is dependent on chain length, extent of unsaturation and pH. There is a requirement for an unesterified carboxyl group. In respiring mitochondria fatty acids act in the presence of nucleotide by lowering the ‘break‐point’ potential at which the conductance of the 32000‐M_r protein increases. Fatty acids have no effect on the chloride uniport activity of the 32000‐M_r protein, but decouple the interference between chloride and protons when the simultaneous transport of both ions is attempted.

In mammalian brown adipose tissue, thermogenesis is explained by uncoupling mitochondrial respiration from ATP synthesis. Uncoupling protein‐1 (UCP1) is responsible for this uncoupled state, because it allows proton re‐entry into the matrix and thus dissipates the proton gradient generated by the respiratory chain. Proton transport by UCP1 is regulated negatively by nucleotides and positively by fatty acids. Adrenergic stimulation of brown adipocytes stimulates lipolysis and therefore enhances uncoupling and thermogenesis. Adrenergic stimulation also boosts ucp1 gene transcription. Since retinoic acid also promotes ucp1 gene transcription and its structure makes it a possible activator of UCP1, we hypothesized that retinoic acid, like noradrenaline, could have a dual action and trigger the activity of the protein UCP1 itself. Here we show that retinoic acid strongly increases proton transport by UCP1 in brown adipose tissue mitochondria and that it is much more potent than fatty acids. These data are corroborated with yeast mitochondria where UCP1 was introduced by genetic manipulation. The yeast expression system allows the comparison of the UCP1 with the newly described homologues UCP2 and UCP3. The search for regulators of UCP2 has demonstrated that it is positively regulated by retinoids in a pH‐dependent manner.

Mouse sperm produce enough ATP to sustain motility by anaerobic glycolysis and respiration. However, previous studies indicated that an active glycolytic pathway is required to achieve normal sperm function, and identified glycolysis as the main source of ATP to fuel the motility of mouse sperm. All the available evidence has been gathered for the lab mouse, while comparative studies including closely related mouse species have revealed (a) a wide range of variation in sperm motility and ATP production, and (b) that the lab mouse shows comparatively low values in these traits. In this study, we compared the relative reliance on the usage of glycolysis or oxidative phosphorylation as ATP sources for sperm motility between mouse species that exhibit significantly different sperm performance parameters. We found that the sperm of species with higher oxygen consumption/lactate excretion rate ratio were able to produce higher amounts of ATP, achieving higher swimming velocities. Additionally, we show that the species with higher respiration/glycolysis ratio have a higher degree of dependence upon active oxidative phosphorylation. Moreover, we characterize for the first time, two mouse species in which sperm depend on functional oxidative phosphorylation to achieve normal performance. Finally, we discuss that sexual selection through sperm competition could promote adaptations in sperm energetic metabolism tending to increase the usage of oxidative phosphorylation in relation to glycolysis, as this would constitute a more efficient pathway for the generation of ATP (and faster sperm).