Hibernating astronauts-science or fiction?

For long-duration manned space missions to Mars and beyond, reduction of astronaut metabolism by torpor, the metabolic state during hibernation of animals, would be a game changer: Water and food intake could be reduced by up to 75% and thus reducing payload of the spacecraft. Metabolic rate reduction in natural torpor is linked to profound changes in biochemical processes, i.e., shift from glycolysis to lipolysis and ketone utilization, intensive but reversible alterations in organs like the brain and kidney, and in heart rate control via Ca2+. This state would prevent degenerative processes due to organ disuse and increase resistance against radiation defects. Neuro-endocrine factors have been identified as main targets to induce torpor although the exact mechanisms are not known yet. The widespread occurrence of torpor in mammals and examples of human hypometabolic states support the idea of human torpor and its beneficial applications in medicine and space exploration.

Phylogenetic differences of mammalian basal metabolic rate are not explained by mitochondrial basal proton leak.

Metabolic rates of mammals presumably increased during the evolution of endothermy, but molecular and cellular mechanisms underlying basal metabolic rate (BMR) are still not understood. It has been established that mitochondrial basal proton leak contributes significantly to BMR. Comparative studies among a diversity of eutherian mammals showed that BMR correlates with body mass and proton leak. Here, we studied BMR and mitochondrial basal proton leak in liver of various marsupial species. Surprisingly, we found that the mitochondrial proton leak was greater in marsupials than in eutherians, although marsupials have lower BMRs. To verify our finding, we kept similar-sized individuals of a marsupial opossum (Monodelphis domestica) and a eutherian rodent (Mesocricetus auratus) species under identical conditions, and directly compared BMR and basal proton leak. We confirmed an approximately 40 per cent lower mass specific BMR in the opossum although its proton leak was significantly higher (approx. 60%). We demonstrate that the increase in BMR during eutherian evolution is not based on a general increase in the mitochondrial proton leak, although there is a similar allometric relationship of proton leak and BMR within mammalian groups. The difference in proton leak between endothermic groups may assist in elucidating distinct metabolic and habitat requirements that have evolved during mammalian divergence.

Photoperiodic control and effects of melatonin on nonshivering thermogenesis and brown adipose tissue.

Exposure to a short photoperiod improved the thermogenic capacity, and cold resistance of Djungarian hamsters and increased the respiratory power of their brown adipose tissue. Exposure to a long photoperiod caused a decrease in thermogenic measurements. This thermotropic action of the short photoperiod was detectable only during late summer and fall. A similar thermotropic response could be elicited by implanting hamsters with melatonin, indicating that the pineal may be involved in photoperiodic control of thermoregulatory effectors.

Marsupial uncoupling protein 1 sheds light on the evolution of mammalian nonshivering thermogenesis.

Brown adipose tissue expressing uncoupling protein 1 (UCP1) is responsible for adaptive nonshivering thermogenesis giving eutherian mammals crucial advantage to survive the cold. The emergence of this thermogenic organ during mammalian evolution remained unknown as the identification of UCP1 in marsupials failed so far. Here, we unequivocally identify the marsupial UCP1 ortholog in a genomic library of Monodelphis domestica. In South American and Australian marsupials, UCP1 is exclusively expressed in distinct adipose tissue sites and appears to be recruited by cold exposure in the smallest species under investigation (Sminthopsis crassicaudata). Our data suggest that an archetypal brown adipose tissue was present at least 150 million yr ago allowing early mammals to produce endogenous heat in the cold, without dependence on shivering and locomotor activity.

Seasonal Control of Mammalian Energy Balance: Recent Advances in the Understanding of Daily Torpor and Hibernation.

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.

Physiological approach to maturation of brown adipocytes in primary cell culture.

Molecular and metabolic aspects of differentiation of brown adipocytes of the Djungarian hamster (Phodopus sungorus) were studied in primary culture. Expression of uncoupling protein and lipoprotein lipase were investigated by Western and Northern blotting and indirect immuno-fluorescence microscopy. The activity of 5'-deiodinase type II was determined by a radioactive enzyme assay. Activity of cytochrome-c-oxidase and cell respiration rates were measured with a Clark electrode. We evaluated functional differences of developmental stages by measuring the reaction to beta-adrenergic stimulation throughout the differentiation process. The results show that differentiation of hamster brown adipocytes is an at least two-step development with physiologically discriminable cell types. Generation of triiodothyronine (T3) from thyroxine by activation of the 5'deiodinase occurs in immature brown adipocytes and is mediated primarily by beta1- rather than beta3-adrenergic receptors. The thermogenic capacity is subsequently increased in mature brown adipocytes. beta-Adrenergic receptor stimulation increases UCP expression of mature adipocytes but is not able to recruit new brown adipocytes.

Contrasting adrenergic effects on lipoprotein lipase gene expression in the brown adipose tissue of intact mice and in cultured brown adipocytes from mice.

To examine the regulation of lipoprotein lipase (LPL) gene expression, LPL mRNA levels in the brown adipose tissue of intact mice and in mouse brown adipocyte cultures were examined. In intact mice, exposure to cold resulted in a rapid, transient, 5-fold increase in LPL mRNA level. Norepinephrine (NE) injection could fully mimic the effect of acute exposure to cold, and LPL mRNA and enzymatic activity were increased in parallel after NE injection. These results indicated positive adrenergic control of LPL gene expression in the brown adipose tissue of intact mice. In cultured mouse brown adipocytes, the level of spontaneously expressed LPL mRNA decreased in parallel with the progression of brown adipocyte differentiation. NE treatment of undifferentiated cells led to a decrease in LPL mRNA levels. In brown adipocytes that had reached a mature state, NE had a small negative or no effect on LPL mRNA levels, irrespective of whether the experiment was performed in the presence or absence of insulin or of newborn-calf serum. It was concluded that LPL gene expression in brown adipose tissue in intact mice is under adrenergic control but that this gene is not under positive adrenergic control in cultured brown adipocytes from mice, although these cells are otherwise adrenergically sensitive. The presence of additional factors may be necessary to confer adrenergic sensitivity to the LPL gene in the cultured brown adipocytes; alternatively, cells other than the mature brown adipocytes may confer the positive adrenergic sensitivity to the brown adipose tissue depots in situ.

Photoperiod and thermoregulation in vertebrates: body temperature rhythms and thermogenic acclimation.

Evidence has recently begun to accumulate that photoperiodic responses of mammals and birds may affect the control of energy balance and thermoregulation. Exposure to short photoperiod can lower the set point for body temperature regulation in birds and mammals, as well as the voluntarily selected body temperature in ectothermic lizards. This decrease is accompanied by a reorganization of circadian or ultradian rhythms of body temperature, particularly an increase in periods spent at rest with minimum body temperatures. Short photoperiod is also used as an environmental cue for induction of seasonal torpor or facilitation of hibernation. During winter, cold tolerance of small mammals is improved by an increase of nonshivering thermogenesis in brown fat. Thermogenic capacity of brown fat (respiratory enzymes, mitochondria, uncoupling protein) is enhanced in response to short photoperiod. This response is mediated via an increase in the activity of sympathetic innervation in brown fat. Moreover, an exposure to short photoperiod prior to low temperatures may act in preparing brown fat for facilitated thermogenesis during acclimation to cold. This shows that photoperiodic control not only affects energy balance indirectly via the control of reproduction or body mass, but may directly interact with central control of thermoregulation and may influence the process of acclimatization.

Inhibition of 5'-deiodination of thyroxine suppresses the cold-induced increase in brown adipose tissue messenger ribonucleic acid for mitochondrial uncoupling protein without influencing lipoprotein lipase activity.

Young adult male and female Djungarian hamsters were exposed to ambient temperatures of 23 or 0 C for 12 h; half of the animals in each group were treated with iopanoic acid to suppress the peripheral conversion of T4 to the thermotropically active thyroid hormone T3 by the enzyme 5'-deiodinase (5'D). Brown adipose tissue (BAT) mRNA for uncoupling protein (UCP), BAT lipoprotein lipase (LPL) activity, and 5'D activity were measured at the conclusion of the study. A temperature of 0 C produced large rises in 5'D and LPL activities and a similar large increase in UCP mRNA within the 12-h exposure period. When 5'D activity was inhibited with iopanoic acid, mRNA for UCP was reduced, while LPL activity was unaffected. The results show that the optimal production of mRNA for BAT UCP depends on the availability of T3; however, T3 is not required for the cold-induced activation of LPL activity in BAT.

Short photoperiod reduces leptin gene expression in white and brown adipose tissue of Djungarian hamsters.

Leptin gene expression in white (WAT) and brown adipose tissue (BAT) of the Djungarian hamster (Phodopus sungorus) was analyzed during seasonal acclimatization. Leptin gene expression in WAT was markedly reduced during winter, independent of changes in environmental temperature. Exposure to artificial short photoperiod also decreased leptin gene expression in WATas well as in BAT. Although specific leptin gene expression was lower in BAT, total depot expression was as high as in WAT depots, due to higher RNA content of BAT. Our results indicate that there is significant leptin synthesis in brown fat and that leptin might be involved in photoperiod mediated seasonal adaptations of mammals independent of food deprivation or overfeeding.

How photoperiod influences body temperature selection in Lacerta viridis.

European green lizards, Lacerta viridis, show a distinct annual cycle in their day and nighttime selected body temperature (T b) levels when monitored under natural photoperiod. The amplitude between daily photophase and scotophase temperatures varies throughout the year. Highest body temperatures with smallest day/night variation are selected from May through July. Throughout fall, the difference between day and nighttime selected T b levels increases. Lizards inevitably enter a state of winter dormancy which terminates daily rhythmicity patterns. Under natural photoperiodic conditions, cessation of dormancy occurs spontaneously by mid-March, regardless whether high temperatures are available or not. Lacerta viridis respond to an artificial long photoperiod (16 h light, 8 h dark) at all times of the year with modifications in both diel patterns and levels of selected T b to summer-like conditions. When, however, the natural photoperiod at different phases in the annual cycle is held constant for six to eight weeks, T b selection of Lacerta viridis also remains stable at the level corresponding to the prevailing photoperiod. These results implicate that the photoperiod is a more prominent Zeitgeber for seasonal cueing of temperature selection than has been surmised in the past. Further, we suggest that the large variations recorded in daily T b cycles do not imply that this lizard is an "imprecise" thermoregulator, but rather indicates an important integral process necessary for seasonal acclimatization.

The seasonal cycle of body weight in the Djungarian hamster: photoperiodic control and the influence of starvation and melatonin.

Photoperiod is the primary environmental cue for seasonality in the Djungarian hamster (Phodopus sungorus). This species reduces its body weight by 30% during winter to reduce its energy requirements. The aim of this study was (1) to examine the influence of food intake on the body weight cycle and (2) to evaluate the role of photoperiod and melatonin in determining this seasonal cycle.While body weight increased slightly (7%) from August through December in long photoperiod, it dropped significantly (32%) in the natural photoperiod group and food intake gradually decreased in this group by about 20%. After two periods of 40% food restriction, the hamsters in long photoperiod rapidly returned to their initial body weight. In contrast, hamsters under natural photoperiod only gained weight until they had reached the body weight of the controls at that particular time.In a second experiment, hamsters were kept under natural photoperiod throughout the year. At 6 different intervals two groups of hamsters were implanted with melatonin capsules and transferred to either short (LD 8:16) or long photoperiod (LD 16:8). The results show that the seasonal change in body weight is caused by a combined action of photoperiod and a change in the animals' sensitivity towards photoperiod and melatonin.

Telemetric field studies of body temperature and activity rhythms of Acomys russatus and A. cahirinus in the Judean Desert of Israel.

Two species of the genus Acomys coexist in arid zones of southern Israel. Acomys russatus is distributed in extremely arid areas, while A. cahirinus is common in both Mediterranean and arid regions. Individuals of both species from a rodent community in the Ein Gedi Nature Reserve were implanted with temperature-sensitive transmitters. Body temperature (T b) rhythms were recorded in free-ranging mice at four different seasons of the year. A. cahirinus (30-45 g) showed a nocturnal rhythm of T b throughout the year. In the activity phase during the night T b increased to 38.2°C. During the day T b decreased to 34°C. This species displayed this pattern in summer also when ambient temperatures rose above T b. The T b of A. russatus (45-65 g) varied between 34.8 and 41°C during the hot season, showing a bimodal temperature rhythm with maximal values in the morning and in the evening. Measurements of activity in this species showed inactivity during the hottest period of a summer day. In winter A. russatus showed no clearly detectable diurnal or ultradian rhythm in T b, which remained constant between narrow limits of 35.2 and 36.8°C.

Seasonal thermogenic acclimation of diurnally and nocturnally active desert spiny mice.

Diurnally active golden spiny mice (Acomys russatus) and nocturnal common spiny mice (Acomys cahirinus) coexist in hot rocky deserts of Israel. Diurnal and nocturnal activities expose these species to different climatic conditions. Nonshivering thermogenesis (NST) capacity of individuals of both species immediately upon removal from the field exhibited seasonal changes, with no significant interspecific difference. Colony-reared mice of either species transferred in the laboratory from long to short photoperiod increased NST capacity, though to a lesser extent than observed in the seasonal acclimatization. The underlying biochemical mechanisms of short photoperiod acclimation differed between the species. In both Cytochrome-c oxidase (Cox) activity was higher in short as compared to long photoperiod. In short-photoperiod-acclimated A. cahirinus uncoupling protein (UCP) content in brown adipose tissue (BAT) was significantly higher than in long photoperiod, while in A. russatus there was no significant change. In A. russatus there was a significant increase in lipoprotein lipase (LPL) activity in BAT in short-photoperiod-acclimated individuals, while in A. cahirinus LPL activity was high under both acclimations. The low LPL activity in brown adipose tissue of desert-adapted A. russatus may facilitate lipid uptake in white adipose tissue, an advantage in desert conditions where food is scarce and irregularly distributed in space and time.

Computerized body temperature telemetry in small animals: use of simple equipment and advanced noise suppression.

Telemetric measurement of body temperatures in small animals implanted with commercially available transmitters (Mini-Mitter) has been automated using the Commodore C-64 microcomputer. Transmitter output frequencies are received by common AM-radios, converted to TTL level by a simple interface and measured by a machine language subroutine. Multiplexed input is under software control and allows for the recording of at least 16 animals selectively, with data points for each sample every 3 min. The effects of environmental electrical noise are significantly reduced by an error detection system based on the separate reception of interference.

Temporal partitioning among diurnally and nocturnally active desert spiny mice: energy and water turnover costs.

Nocturnal Acomys cahirinus and diurnally active A. russatus coexist in hot rocky deserts. Diurnal and nocturnal activity exposes them to different climatic challenges. A doubly-labelled water field study revealed no significant differences in water turnover between the species at all seasons, reflecting the adaptations of A. russatus to water conservation. In summers the energy expenditure of A. russatus tended to be higher than that of A. cahirinus. Energy requirements of A. cahirinus in winter are double than that of A. russatus, and may reflect the cost of thermoregulating during cold nights.

Photoperiod-dependent regulation of carboxypeptidase E affects the selective processing of neuropeptides in the seasonal Siberian hamster (Phodopus sungorus).

The production of bioactive peptides from biologically inactive precursors involves extensive post-translational processing, including enzymatic cleavage by proteolytic peptidases. Endoproteolytic prohormone-convertases initially cleave the precursors of many neuropeptides at specific amino acid sequences to generate intermediates with basic amino acid extensions on their C-termini. Subsequently, the related exopeptidases, carboxypeptidases D and E (CPD and CPE), are responsible for removing these amino acids before the peptides achieve biological activity. We investigated the effect of photoperiod on the processing of the neuropeptide precursor pro-opiomelanocortin (POMC) and its derived neuropeptides, α-melanocyte-stimulating hormone (MSH) and ß-endorphin (END), within the hypothalamus of the seasonal Siberian hamster (Phodopus sungorus). We thus compared hypothalamic distribution of CPD, CPE, α-MSH and ß-END using immunohistochemistry and measured the enzyme activity of CPE and concentrations of C-terminally cleaved α-MSH in short-day (SD; 8 : 16 h light/dark) and long-day (LD; 16 : 8 h light/dark) acclimatised hamsters. Increased immunoreactivity (-IR) of CPE, as well as higher CPE activity, was observed in SD. This increase was accompanied by more ß-END-IR cells and substantially higher levels of C- terminally cleaved α-MSH, as determined by radioimmunoassay. Our results suggest that exoproteolytic cleavage of POMC-derived neuropeptides is tightly regulated by photoperiod in the Siberian hamster. Higher levels of biological active α-MSH- and ß-END in SD are consistent with the hypothesis that post-translational processing is a key event in the regulation of seasonal energy balance.

Torpor patterns, arousal rates, and temporal organization of torpor entry in wildtype and UCP1-ablated mice.

In eutherian mammals, uncoupling protein 1 (UCP1) mediated non-shivering thermogenesis from brown adipose tissue (BAT) provides a mechanism through which arousal from torpor and hibernation is facilitated. In order to directly assess the magnitude by which the presence or absence of UCP1 affects torpor patterns, rewarming and arousal rates within one species we compared fasting induced torpor in wildtype (UCP1(+/+)) and UCP1-ablated mice (UCP(-/-)). Torpor was induced by depriving mice of food for up to 48 h and by a reduction of ambient temperature (T (a)) from 30 to 18°C at four different time points after 18, 24, 30 and 36 h of food deprivation. In most cases, torpor bouts occurred within 20 min after the switch in ambient temperature (30-18°C). Torpor bouts expressed during the light phase lasted 3-6 h while significantly longer bouts (up to 16 h) were observed when mice entered torpor during the dark phase. The degree of hypometabolism (5-22 ml h(-1)) and hypothermia (19.5-26.7°C) was comparable in wildtype and UCP1-ablated mice, and both genotypes were able to regain normothermia. In contrast to wildtype mice, UCP1-ablated mice did not display multiple torpor bouts per day and their peak rewarming rates from torpor were reduced by 50% (UCP1(+/+): 0.24 ± 0.08°C min(-1); UCP1(-/-): 0.12 ± 0.04°C min(-1)). UCP1-ablated mice therefore took significantly longer to rewarm from 25 to 32°C (39 vs. 70 min) and required 60% more energy for this process. Our results demonstrate the energetic benefit of functional BAT for rapid arousal from torpor. They also suggest that torpor entry and maintenance may be dependent on endogenous rhythms.

Seasonal changes of myostatin expression and its relation to body Mass acclimation in the Djungarian hamster, Phodopus sungorus.

The Djungarian hamster is an animal that is prominent for distinct seasonal adaptations. Cued by shortening day length in autumn they spontaneously exhibit reductions in food intake, body mass (BM), fat mass and also in lean mass (LM). The mechanisms behind the seasonal regulation of body composition are only partly resolved. Although most studies focused on the participation of body fat in seasonal body weight regulation, we addressed the influence of LM, moreover of muscle mass (MM) on seasonal BM changes. Therefore, we analyzed body composition, MM and the expression of myostatin, a hormone negatively regulating muscle growth and differentiation, in Djungarian hamsters in response to naturally changing photoperiod in winter compared to long photoperiod (LP). Winter-acclimated hamsters upregulated myostatin mRNA when compared with hamsters adapted to natural and artificial LP, whereas MM remained unchanged when compared with natural LP. Moreover, in natural short photoperiod, individual myostatin expression levels were negatively correlated with MM. These results suggest that myostatin is under seasonal control in order to regulate MM and hence contributes to the overall LM and therefore BM changes in seasonal mammals.