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Endothermic mammals and birds require intensive energy turnover to sustain high body temperatures and metabolic rates. To cope with the energetic bottlenecks associated with the change of seasons, and to minimise energy expenditure, complex mechanisms and strategies are used, such as daily torpor and hibernation. During torpor, metabolic depression and low body temperatures save energy. However, these bouts of torpor, lasting for hours to weeks, are interrupted by active 'euthermic' phases with high body temperatures. These dynamic transitions require precise communication between the brain and peripheral tissues to defend rheostasis in energetics, body mass and body temperature. The hypothalamus appears to be the major control centre in the brain, coordinating energy metabolism and body temperature. The sympathetic nervous system controls body temperature by adjustments of shivering and nonshivering thermogenesis, with the latter being primarily executed by brown adipose tissue. Over the last decade, comparative physiologists have put forward integrative studies on the ecophysiology, biochemistry and molecular regulation of energy balance in response to seasonal challenges, food availability and ambient temperature. Mammals coping with such environments comprise excellent model organisms for studying the dynamic regulation of energy metabolism. Beyond the understanding of how animals survive in nature, these studies also uncover general mechanisms of mammalian energy homeostasis. This research will benefit efforts of translational medicine aiming to combat emerging human metabolic disorders. The present review focuses on recent advances in the understanding of energy balance and its neuronal and endocrine control during the most extreme metabolic fluctuations in nature: daily torpor and hibernation.

Mitochondrial membrane potential (ΔΨM) is a central intermediate in oxidative energy metabolism. Although ΔΨM is routinely measured qualitatively or semi-quantitatively using fluorescent probes, its quantitative assay in intact cells has been limited mostly to slow, bulk-scale radioisotope distribution methods. Here we derive and verify a biophysical model of fluorescent potentiometric probe compartmentation and dynamics using a bis-oxonol-type indicator of plasma membrane potential (ΔΨP) and the ΔΨM probe tetramethylrhodamine methyl ester (TMRM) using fluorescence imaging and voltage clamp. Using this model we introduce a purely fluorescence-based quantitative assay to measure absolute values of ΔΨM in millivolts as they vary in time in individual cells in monolayer culture. The ΔΨP-dependent distribution of the probes is modelled by Eyring rate theory. Solutions of the model are used to deconvolute ΔΨP and ΔΨM in time from the probe fluorescence intensities, taking into account their slow, ΔΨP-dependent redistribution and Nernstian behaviour. The calibration accounts for matrix:cell volume ratio, high- and low-affinity binding, activity coefficients, background fluorescence and optical dilution, allowing comparisons of potentials in cells or cell types differing in these properties. In cultured rat cortical neurons, ΔΨM is −139 mV at rest, and is regulated between −108 mV and −158 mV by concerted increases in ATP demand and Ca2+-dependent metabolic activation. Sensitivity analysis showed that the standard error of the mean in the absolute calibrated values of resting ΔΨM including all biological and systematic measurement errors introduced by the calibration parameters is less than 11 mV. Between samples treated in different ways, the typical equivalent error is ∼5 mV.