The matching of energy supply and demand under hypoxic conditions is critical for sustaining myocardial function. Numerous reports indicate that basal energy requirements and ion handling may differ between the ventricles. We hypothesized that ventricular response to hypoxia shows inter-ventricular differences caused by the heterogeneity in glucose metabolism and expression and activity of ion transporters. Thus we assessed glucose utilization rate, ATP, sodium and potassium concentrations, Na,K-ATPase activity and tissue reduced:oxidised glutathione (GSH/GSSG) content in the right and left ventricles before and after the exposure of either the whole animals or isolated blood-perfused hearts to hypoxia. The hypoxia-induced boost in glucose utilization was more pronounced in the left ventricle compared to the right one. ATP levels in the right ventricle of hypoxic heart were lower than those in the left ventricle. Left ventricular sodium content was higher and hydrolytic Na,K-ATPase activity was reduced compared with the right ventricle. Administration of the Na, K-ATPase blocker ouabain caused rapid increase in the right ventricular Na+ and elimination of the inter-ventricular Na(+) gradients. Exposure of the hearts to hypoxia made the inter-ventricular heterogeneity in the Na(+) distribution even more pronounced. Furthermore, systemic hypoxia caused oxidative stress that was more pronounced in the right ventricle as revealed by GSH/GSSG ratios. Based on these findings, we suggest that the right ventricle is more prone to hypoxic damage as it is less efficient in recruiting glucose as an alternative fuel and is particularly dependent on the efficient Na, K-ATPase function.