Advances in molecular biology of hibernation in mammals

Mammalian hibernation is characterized by profound reductions in metabolism, oxygen consumption and heart rate. As a result, the animal enters a state of suspended animation where core body temperatures can plummet as low as -2.9 degrees C. Not only can hibernating mammals survive these physiological extremes, but they also return to a normothermic state of activity without reperfusion injury or other ill effects. This review examines recent findings on the genes, proteins and small molecules that control the induction and maintenance of hibernation in mammals. The molecular events involved with remodeling metabolism, inducing hypothermia and maintaining organ function are discussed and considered with respect to analogous processes in non-hibernating mammals such as mice and humans. The advent of sequenced genomes from three distantly related hibernators, a bat, hedgehog and ground squirrel, provides additional opportunities for molecular biologists to explore the mechanistic aspects of this biological adaptation in greater detail.     

Mitochondrial metabolism in hibernation and daily torpor: a review

Hibernation and daily torpor involve substantial decreases in body temperature and metabolic rate, allowing birds and mammals to cope with cold environments and/or limited food. Regulated suppression of mitochondrial metabolism probably contributes to energy savings: state 3 (phosphorylating) respiration is lower in liver mitochondria isolated from mammals in hibernation or daily torpor compared to normothermic controls, although data on state 4 (non-phosphorylating) respiration are equivocal. However, no suppression is seen in skeletal muscle, and there is little reliable data from other tissues. In both daily torpor and hibernation, liver state 3 substrate oxidation is suppressed, especially upstream of electron transport chain complex IV. In hibernation respiratory suppression is reversed quickly in arousal even when body temperature is very low, implying acute regulatory mechanisms, such as oxaloacetate inhibition of succinate dehydrogenase. Respiratory suppression depends on in vitro assay temperature (no suppression is evident below ~30 degrees C) and (at least in hibernation) dietary polyunsaturated fats, suggesting effects on inner mitochondrial membrane phospholipids. Proton leakiness of the inner mitochondrial membrane does not change in hibernation, but this also depends on dietary polyunsaturates. In contrast proton leak increases in daily torpor, perhaps limiting reactive oxygen species production.     

生长抑素的生理作用及其与哺乳动物冬眠启动的关系

生长抑素(somatostatin, SS)是一种具有抑制性作用的脑肠肽,在哺乳动物的中枢神经系统及消化系统中广泛分布,具有抑制多种垂体激素的分泌、维持胃肠道功能等重要作用。冬眠是一种低体温和低代谢状态,以全身多器官功能暂时"休止"为特征,依据生长抑素特殊的"刹车"作用,推测其与冬眠启动过程相关。本文综述了生长抑素的生理作用及其与哺乳动物冬眠启动的关系。     

Metabolic rate depression and biochemical adaptation in anaerobiosis, hibernation and estivation

For many animals, the best defense against harsh environmental conditions is an escape to a hypometabolic or dormant state. Facultative metabolic rate depression is the common adaptive strategy of anaerobiosis, hibernation, and estivation, as well as a number of other arrested states. By reducing metabolic rate by a factor ranging from 5 to 100 fold or more, animals gain a comparable extension of survival time that can support months or even years of dormancy. The present review focuses on the molecular control mechanisms that regulate and coordinate cellular metabolism for the transition into dormancy. These include reversible control over the activity state of enzymes via protein phosphorylation or dephosphorylation reactions, pathway regulation via the association or dissociation of particle-bound enzyme complexes, and fructose-2,6-bisphosphate regulation of the use of carbohydrate reserves for biosynthetic purposes. These mechanisms, their interactions, and the regulatory signals (e.g., second messenger molecules, pH) that coordinate them form a common molecular basis for metabolic depression in anoxia-tolerant vertebrates (goldfish, turtles) and invertebrates (marine molluscs), hibernation in small mammals, and estivation in land snails and terrestrial toads.     

Retaining memory after hibernation: Performance varies independently of activity levels in wild grey mouse lemurs

Hibernation, a hypometabolic state associated with low body temperature and reduced metabolic and activity rates, represents one adaptation to harsh seasonal environmental conditions. As a consequence of hypometabolism, energetically costly neuronal processes also ought to be reduced. Since active neuronal pathways are prerequisites for learning and memory, and because previous studies revealed variable patterns, it remains unclear whether and how hibernating animals retain memories, however. Here, we investigated the effect of seasonally reduced activity on memory retention in 36 wild grey mouse lemurs (Microcebus murinus). Data from activity loggers confirmed that female grey mouse lemurs entered hibernation during the cool dry season, whereas males exhibited episodic bursts of activity throughout the austral winter. Thus, compared to males, we predicted females to show lower memory retention of visual and spatial stimulus–reward associations learned before hibernation. In contrast to our prediction, all individuals performed worse in the post-hibernation testing session in both types of tests, compared to the pre-hibernation learning session, and males (N = 11) performed even worse than females (N = 14) in the post-hibernation testing session. Although females (N = 9) equipped with activity loggers tended to be less active than males (N = 4), sex-specific activity levels were unrelated to interindividual differences in memory retention. Hence, the post-hibernation decrease in performance of grey mouse lemurs may reflect a more general disability to retain stimulus–reward associations than a lack of memory retention due to seasonal hypometabolism, as suggested for some species of bats or squirrels.     

Human torpor: translating insights from nature into manned deep space expedition

During a long-duration manned spaceflight mission, such as flying to Mars and beyond, all crew members will spend a long period in an independent spacecraft with closed-loop bioregenerative life-support systems. Saving resources and reducing medical risks, particularly in mental heath, are key technology gaps hampering human expedition into deep space. In the 1960s, several scientists proposed that an induced state of suppressed metabolism in humans, which mimics ‘hibernation’, could be an ideal solution to cope with many issues during spaceflight. In recent years, with the introduction of specific methods, it is becoming more feasible to induce an artificial hibernation-like state (synthetic torpor) in non-hibernating species. Natural torpor is a fascinating, yet enigmatic, physiological process in which metabolic rate (MR), body core temperature (T_b) and behavioural activity are reduced to save energy during harsh seasonal conditions. It employs a complex central neural network to orchestrate a homeostatic state of hypometabolism, hypothermia and hypoactivity in response to environmental challenges. The anatomical and functional connections within the central nervous system (CNS) lie at the heart of controlling synthetic torpor. Although progress has been made, the precise mechanisms underlying the active regulation of the torpor–arousal transition, and their profound influence on neural function and behaviour, which are critical concerns for safe and reversible human torpor, remain poorly understood. In this review, we place particular emphasis on elaborating the central nervous mechanism orchestrating the torpor–arousal transition in both non-flying hibernating mammals and non-hibernating species, and aim to provide translational insights into long-duration manned spaceflight. In addition, identifying difficulties and challenges ahead will underscore important concerns in engineering synthetic torpor in humans. We believe that synthetic torpor may not be the only option for manned long-duration spaceflight, but it is the most achievable solution in the foreseeable future. Translating the available knowledge from natural torpor research will not only benefit manned spaceflight, but also many clinical settings attempting to manipulate energy metabolism and neurobehavioural functions.     

Membrane function in mammalian hibernation

For homeotherms the maintenance of a high, uniform body temperature requires a constant energy supply and food intake. For many small mammals, the loss of heat in winter exceeds energy supply, particularly when food is scarce. To survive, some animals have developed a capacity for adaptive hypothermia in which they lower their body temperature to a new regulatory set-point, usually a few degrees above the ambient. This process, generally known as hibernation, reduces the temperature differential, metabolic activity, as well as the energy demand, and thus facilitates survival during winter. Successful hibernation in mammals requires that the enzymatic processes are regulated in such a manner that metabolic balance is maintained at both the high body temperature of the summer-active animal (37 degrees C) and the low body temperature of the winter-torpid animal (approx. 5 degrees C). This means that the cellular membranes have thermal properties capable of maintaining a balanced metabolism at these extreme physiological temperatures. The available evidence indicates that, for some tissues, preparation for hibernation involves an alteration in the lipid composition and thermal properties of cellular membranes. Marked differences in the thermal response of cellular membranes have been observed on a seasonal basis and, in some membranes, differences in lipid composition have been associated with the torpid state. However, to date, no consistent changes in lipid composition which would account for, or explain, the changes in membrane thermal response, have been detected. An important point to emphasize is that the process of 'homeoviscous adaptation', which occurs in procaryotes and some poikilotherms during acclimation to low temperatures, is not a characteristic feature of most membranes of mammalian hibernators.     

Regulation of Akt during torpor in the hibernating ground squirrel, Ictidomys tridecemlineatus

The 13-lined ground squirrel (Ictidomys tridecemlineatus) is capable of entering into extended periods of torpor during winter hibernation. The state of torpor represents a hypometabolic shift wherein the rate of oxygen consuming processes are strongly repressed in an effort to maintain cellular homeostasis as the availability of food energy becomes limited. We are interested in studying hibernation/torpor because of the robust state of tolerance to constrained oxygen delivery, oligemia, and hypothermia achieved by the tissues of hibernating mammals. The role of the serine/threonine kinase Akt (also known as PKB) has been examined in torpor in previous studies. However, this is the first study that examines the level of Akt phosphorylation in the liver during the two transition phases of the hibernation cycle: entrance into torpor, and the subsequent arousal from torpor. Our results indicate that Akt is activated in the squirrel liver by phosphorylation of two key residues (Thr³⁰⁸ and Ser⁴⁷³) during entrance into torpor and arousal from torpor. Moreover, we observed increased phosphorylation of key substrates of Akt during the two transition stages of torpor. Finally, this study reports the novel finding that PRAS40, a component of the TORC1 multi-protein complex and a potentially important modulator of metabolism, is regulated during torpor.     

Hypometabolism in Aotus trivirgatus (Primates, Plathyrhini, Cebidae)]

The standard metabolism of Aotus trivirgatus (Night monkey, Owl monkey) is 22.5 to 46.2 per cent below Kleiber's prevision curve for mammals, which applies to other cebid monkeys like Saimiri sciureus and Alouatta. However the metabolic rate of Aotus is not reduced to the extent found in two hypometabolic prosimians Perodicticus potto and Nycticebus coucang. The low metabolism in Aotus is associated with a normal body temperature and a thick fur of high insulating power. These results are discussed.     

Daily torpor and hibernation in birds and mammals

Many birds and mammals drastically reduce their energy expenditure during times of cold exposure, food shortage, or drought, by temporarily abandoning euthermia, i.e. the maintenance of high body temperatures. Traditionally, two different types of heterothermy, i.e. hypometabolic states associated with low body temperature (torpor), have been distinguished: daily torpor, which lasts less than 24 h and is accompanied by continued foraging, versus hibernation, with torpor bouts lasting consecutive days to several weeks in animals that usually do not forage but rely on energy stores, either food caches or body energy reserves. This classification of torpor types has been challenged, suggesting that these phenotypes may merely represent extremes in a continuum of traits. Here, we investigate whether variables of torpor in 214 species (43 birds and 171 mammals) form a continuum or a bimodal distribution. We use Gaussian‐mixture cluster analysis as well as phylogenetically informed regressions to quantitatively assess the distinction between hibernation and daily torpor and to evaluate the impact of body mass and geographical distribution of species on torpor traits. Cluster analysis clearly confirmed the classical distinction between daily torpor and hibernation. Overall, heterothermic endotherms tend to be small; hibernators are significantly heavier than daily heterotherms and also are distributed at higher average latitudes (～35°) than daily heterotherms (～25°). Variables of torpor for an average 30 g heterotherm differed significantly between daily heterotherms and hibernators. Average maximum torpor bout duration was >30‐fold longer, and mean torpor bout duration >25‐fold longer in hibernators. Mean minimum body temperature differed by ～13°C, and the mean minimum torpor metabolic rate was ～35% of the basal metabolic rate (BMR) in daily heterotherms but only 6% of BMR in hibernators. Consequently, our analysis strongly supports the view that hibernators and daily heterotherms are functionally distinct groups that probably have been subject to disruptive selection. Arguably, the primary physiological difference between daily torpor and hibernation, which leads to a variety of derived further distinct characteristics, is the temporal control of entry into and arousal from torpor, which is governed by the circadian clock in daily heterotherms, but apparently not in hibernators.     

Physiological ecology of cheirogaleid primates: variation in hibernation and torpor

Torpor (i.e. the reduction of body temperature and metabolic rate for less than 24 h) and hibernation (i.e. torpor phases longer than 24 h) are among the most extreme adaptations to seasonality in primate habitats. Although widespread among mammals, this form of extreme thermoregulation is rare among primates and is reported only for species of the cheirogaleid family. Understanding their physiological ecology is crucial for many aspects of cheirogaleid socioecology like their social organization and their mating systems. This paper first provides an overview of published information on hibernation and torpor and identifies a patchy distribution for the occurrence of hibernation across genera, species and populations. Based on a review of published studies from the wild and from captivity, we then propose a possible explanation for variation in hibernation behavior among Microcebus species and populations. Accordingly, the amount of energy that can be saved during torpor early in the lean dry season, which is determined by the minimum ambient temperature will be decisive. Only where temperatures are low, early dry season torpor bouts will be long enough to save enough energy to build up fat reserves for longer bouts of hibernation. Finally, we summarize information on the causal factors for the occurrence of hibernation by analyzing sex differences within populations. Further physiological studies on other cheirogaleid species are needed to identify the phylogenetic origin of hibernation in primates.     

冬眠哺乳动物氧化应激产生及抗氧化防御机制研究进展

哺乳动物的冬眠是一种季节性异温状态,是对外界恶劣自然环境的一种适应策略。冬眠-阵间觉醒周期中,伴随着生理功能的剧烈变化,从冬眠期间整体代谢的抑制,到阵间觉醒时氧代谢的急剧增加,使动物体内产生了大量的氧自由基。然而,冬眠动物出眠时并未表现出明显的氧化损伤迹象,因此,冬眠哺乳动物被认为是一种天然的抗氧化损伤模型。本文从氧化应激的产生、活性氧的来源、抗氧化防御等方面综述了冬眠哺乳动物对氧化应激的防御,并从其抗氧化的分子调控方面分析了冬眠哺乳动物对氧化应激的适应机制。     

Regulation of cardiac rhythm in hibernating mammals

The dramatic fall in heart rate exhibited by mammals entering hibernation begins before there is any noticeable fall in body temperature. The initial, progressive decrease in heart rate is the result of a cyclic parasympathetic activation that induces skipped beats and regular asystoles as well as slows the even heart beat. As body temperature subsequently falls, the parasympathetic influence is progressively withdrawn and periods of parasympathetic and sympathetic dominance alternate and give rise to regular periods of arrhythmia (tachycardia followed by bradycardia), and occasional long asystoles or periods of highly irregular cardiac activity. Superimposed on this is a vagally-mediated, respiratory sinus arrhythmia that is accentuated in species that breathe episodically. These events give way to a uniform heart rate in deep hibernation at low temperatures where both parasympathetic and sympathetic tone appear absent. The complete absence of tone is not a function of reduced temperature but is reflective of the state of deep, steady state hibernation. The elevation in heart rate that accompanies the onset of arousal is the result of dramatic increases in sympathetic activation that precede any increases in body temperature. As body temperature then rises, sympathetic influence is slowly withdrawn. Arrhythmias are also common during natural arousals or shifts from lower to warmer hibernation temperatures as periods of parasympathetic and sympathetic dominance again alternate en route to re-establishing a steady state in euthermia. The mechanism behind, and the biological significance of, cardiac changes mediated through orchestrated arrhythmias remain unknown.     

Depression of mitochondrial respiration during daily torpor of the Djungarian hamster,Phodopus sungorus,is specific for liver and correlates with body temperature

Small mammals actively decrease metabolism during daily torpor and hibernation to save energy. Increasing evidence suggests depression of mitochondrial respiration during daily torpor of the Djungarian hamster but tissue-specificity and relation to torpor depth is unknown. We first confirmed a previous study by Brown and colleagues reporting on the depressed substrate oxidation in isolated liver mitochondria of the Djungarian hamster (Phodopus sungorus) during daily torpor. Next, we show that mitochondrial respiration is not depressed in kidneys, skeletal muscle and heart. In liver mitochondria, we found that state 3 and state 4 respirations correlate with body temperature, suggesting inhibition related to torpor depth and to metabolic rate. We conclude that molecular events leading to depression of mitochondrial respiration during daily torpor are specific to liver and linked to a decrease in body temperature. Different tissue-specificity of mitochondrial depression may assist to compare and identify the molecular nature of mitochondrial alterations during torpor.     

中国哺乳动物生理生态学研究进展与展望

中国哺乳动物生理生态学研究自20世纪50年代始,经过70多年的发展,已系统研究了分布于青藏高原、内蒙古草原、横断山脉等地理分布区的代表性物种的生理适应性,研究主题包括能量代谢和体温调节、冬眠（蛰眠）、水代谢、生态免疫、肠道菌群与宿主的能量代谢和产热调节,研究物种以小型哺乳动物为主。在新时期除了进一步加强对极端环境的生理适应研究外,也需关注大型动物对环境的生理适应,发展新兴领域如保护生理学等,同时要借助多组学技术、同位素技术、遥感技术、红外技术等,加强对动物生理适应的机理性探究。本文回顾了中国哺乳动物生理生态学的发展历程,总结了主要领域取得的重要进展。     

A phenology of the evolution of endothermy in birds and mammals

Recent palaeontological data and novel physiological hypotheses now allow a timescaled reconstruction of the evolution of endothermy in birds and mammals. A three‐phase iterative model describing how endothermy evolved from Permian ectothermic ancestors is presented. In Phase One I propose that the elevation of endothermy – increased metabolism and body temperature (T_b) – complemented large‐body‐size homeothermy during the Permian and Triassic in response to the fitness benefits of enhanced embryo development (parental care) and the activity demands of conquering dry land. I propose that Phase Two commenced in the Late Triassic and Jurassic and was marked by extreme body‐size miniaturization, the evolution of enhanced body insulation (fur and feathers), increased brain size, thermoregulatory control, and increased ecomorphological diversity. I suggest that Phase Three occurred during the Cretaceous and Cenozoic and involved endothermic pulses associated with the evolution of muscle‐powered flapping flight in birds, terrestrial cursoriality in mammals, and climate adaptation in response to Late Cenozoic cooling in both birds and mammals. Although the triphasic model argues for an iterative evolution of endothermy in pulses throughout the Mesozoic and Cenozoic, it is also argued that endothermy was potentially abandoned at any time that a bird or mammal did not rely upon its thermal benefits for parental care or breeding success. The abandonment would have taken the form of either hibernation or daily torpor as observed in extant endotherms. Thus torpor and hibernation are argued to be as ancient as the origins of endothermy itself, a plesiomorphic characteristic observed today in many small birds and mammals.     

Testing the flexibility of fasting-induced hypometabolism in birds: effect of photoperiod and repeated food deprivations

We tested whether photoperiod and repeated fasts modulate the fasting-induced nocturnal hypometabolic response in Japanese quails and pigeons. Quails did not compensate a shorter dark phase by a deeper hypothermia, but instead showed diurnal hypothermia that seemed to be conditioned by the removal of food. Body mass loss measurements indicated that this response saved energy. Repeated fasts in constant photoperiod in quails and pigeons slightly augmented the hypometabolic response only in quails. During fasting intervals, body mass increased in pigeons, but decreased in quails. Apart from the diurnal hypothermia seen in quails, photoperiod and repeated fasts had little influence on fasting-induced hypometabolism.