Extensive use of torpor in 13-lined ground squirrels in the fall prior to cold exposure

Mammalian hibernation is characterized by profound reductions in body temperature (T _b) and metabolic, heart and respiratory rates. These reductions are characteristic of torpor, which is temporally confined to winter. Hibernators including ground squirrels are heterothermic in winter, cycling between multiday periods of torpor with low T _b and brief periods of rewarming. In contrast, ground squirrels remain homeothermic during summer, like non-hibernating mammals. The transition between the homeothermic and heterothermic phases of the circannual rhythm of hibernation is often overlooked in hibernation studies. Here, we examined the use of torpor throughout the fall transition in laboratory-housed 13-lined ground squirrels by recording core body temperature with an implanted data logger. As is typical of laboratory-based hibernation studies, animals were kept in standard housing prior to being moved into a cold, dark room to simulate natural hibernation conditions. Significantly, the vast majority of both male and female ground squirrels expressed torpor in the fall while still housed conventionally and prior to cold exposure. The expression of torpor was not predicted by body weight or age, rather it appears to be preprogrammed in a time-dependent manner that is independent of, yet enhanced by, environmental cues. The timing and duration of these torpor bouts occurring prior to cold exposure were also remarkably sporadic. Thus, it is not possible to know with certainty which animals are torpor-naive before cold exposure in the absence of continuous measurement of body temperature. We conclude that fall animals encompass variable points in the transition between summer and winter phases of the circannual cycle of hibernation, thereby confounding studies in which they are used as non-hibernating controls. Conversely, these fall transition animals offer unique opportunities to define the molecular changes that accompany and enable hibernation.     

The effects of caloric restriction on the body composition and hibernation of the golden-mantled ground squirrel (Spermophilus lateralis)

In preparation for hibernation, golden-mantled ground squirrels (Spermophilus lateralis) must deposit sufficient amounts of lipid during the summer to survive winter hibernation. We conducted an experiment from May 1998 to February 1999 to examine the effects of caloric restriction on the body composition (lipid and fat-free mass) and hibernation of golden-mantled ground squirrels. Ground squirrels were either provided with food ad lib. (controls) or with only enough food to maintain a constant body mass throughout the experiment (calorically restricted). Changes in body composition were followed using total body electrical conductivity (TOBEC). Implanted data loggers that recorded body temperature were used to determine when ground squirrels entered their first torpor bout and the lengths of torpor bouts. Body composition did not change in the calorically restricted ground squirrels between May and September, while both lipid and fat-free mass increased in the controls. However, from September to February, calorically restricted ground squirrels lost only fat-free mass, not lipid mass, but controls lost both lipid and fat-free mass. Calorically restricted ground squirrels entered their first torpor bout about 4 wk after controls, but the torpor bout duration (or length) during hibernation did not differ between the two groups. These results show that ground squirrels maintain body composition during caloric restriction, and the limited quantities of stored lipid have an effect on when hibernation begins but not on torpor bout length.     

Daily rhythmicity and hibernation in the Anatolian ground squirrel under natural and laboratory conditions

We studied daily rhythmicity of body temperature (T _b) before and during hibernation in Anatolian ground squirrels (Spermophilus xanthoprymnus) under natural and laboratory conditions using surgically implanted temperature loggers. Under both conditions, robust daily T _b rhythmicity with parameters comparable to those of other ground squirrel species was observed before but not during hibernation. Euthermic animals had robust daily T _b rhythms with a mean of 37.0°C and a range of excursion of approximately 4°C. No T _b rhythm was detected during torpor bouts, either because T _b rhythmicity was absent or because the daily range of excursion was smaller than 0.2°C. The general patterns of hibernation that we observed in Anatolian ground squirrels were similar to those previously observed by other investigators in other species of ground squirrels.     

Circannual cycles in golden-mantled ground squirrels: Experiments with food deprivation and effects of temperature on periodicity

Summary Female golden-mantled ground squirrels,Spermophilus lateralis, kept in a cool room (9.5 °C), were food deprived and their hibernation thereby extended by 11 weeks on average compared to undeprived control animals in the same room. The deprived group was then refed. Despite their prolonged hibernation, peak weights and onset of hibernation in the subsequent season were only 2–4 weeks later than in the undeprived controls and this effect was only marginally significant. However, the periods of circannual cycles of both the deprived and undeprived groups in the cool room were significantly longer than those of another group of female ground squirrels kept in a warm room (21 °C). It is argued that warm temperatures are capable of terminating the winter phase of circannual cycles, namely that of hibernation and declining body weight. If animals are kept continuously in a cool environment this phase ends spontaneously. Even many weeks of food deprivation are only capable of extending this phase by a relatively small amount, if at all.The author thanks Kirsteen Lang for invaluable help at every phase of this work, Linda Jones and others at the Environmental Sciences Centre, Kananaskis, for heir cooperation, and Janice Farintosh, John Ting and Yasmin Shah for collecting data. Support came from the Natural Sciences and Engineering Research Council of Canada.     

光照黑暗循环条件下达乌尔黄鼠的冬眠模式和能量消耗

为研究冬眠季节的光照条件对贮脂类冬眠动物入眠的影响,在达乌尔黄鼠腹腔埋植体温记录元件iButton,在体重高峰后的下降阶段置于5℃和12L:12D的光照条件下,观察测定其冬眠模式和能量消耗。达乌尔黄鼠冬眠模式出现深冬眠型、少冬眠型和不冬眠型,蛰眠阵包括深冬眠阵、短冬眠阵和日眠阵。不同冬眠阵中最低体温、冬眠阵的持续时间、阵间产热的持续时间、冷却速率和复温速率差异显著;阵间产热的最高体温基本相同。平均每日能量消耗在日眠阵中最高、短冬眠阵中居中、深冬眠阵中最低。入眠时间多集中于黑暗时相,觉醒时间多集中于光照时相。本实验结果提示,在冬眠季节施加光照黑暗循环条件可减少达乌尔黄鼠冬眠的时间,升高阵间最低体温,缩短冬眠阵与阵间产热的持续时间,降低复温速率;增加冬眠期间能量消耗。入眠与觉醒受光照条件影响,具有明显的光暗节律。     

Nutrient constraints in the feeding ecology of an omnivore in a seasonal environment

Summary Nutrient requirements of adult, nonreproductive, omnivorous antelope ground squirrels (Ammospermophilus leucurus) were compared with the nutritional value of their food resources. It was found that nutrient constraints would be important factors in ground squirrel feeding ecology primarily in winter. Potentially important constraints were the requirement for water and nitrogen, and a digestive requirement that average dry matter digestibility of the diet exceed ca. 50%. An unlikely constraint was the requirement for any specific mineral. A linear programming model was used to determine potential diets ground squirrels could consume which satisfied these nutritional requirements and also the ground squirrel's daily energy requirements. During spring ground squirrels could be strict herbivores, but during winter before winter rains ground squirrels had to eat some arthropods to satisfy water requirements.These ground squirrels are not energy maximizers because they spend only one third of their activity period feeding and do not accumulate excess energy as fat. Thus, optimum diets were predicted for winter and spring assuming the goal of feeding time minimization. The model correctly predicted that in wintertime ground squirrels would be primarily granivorous but would consume about 20% arthropods, and that they would switch to herbivory in springtime. Ground squirrels, however, selected a wider dietary range than predicted in both winter and spring. Possible reasons for this discrepancy include an inappropriate assumption that ground squirrels forage for food classes nonsimultaneously, and the possibility that ground squirrels employ sampling as part of their foraging behavior.     

Data logging of body temperatures provides precise information on phenology of reproductive events in a free-living arctic hibernator

Precise measures of phenology are critical to understanding how animals organize their annual cycles and how individuals and populations respond to climate-induced changes in physical and ecological stressors. We show that patterns of core body temperature (T _b) can be used to precisely determine the timing of key seasonal events including hibernation, mating and parturition, and immergence and emergence from the hibernacula in free-living arctic ground squirrels (Urocitellus parryii). Using temperature loggers that recorded T _b every 20 min for up to 18 months, we monitored core T _b from three females that subsequently gave birth in captivity and from 66 female and 57 male ground squirrels free-living in the northern foothills of the Brooks Range Alaska. In addition, dates of emergence from hibernation were visually confirmed for four free-living male squirrels. Average T _b in captive females decreased by 0.5–1.0°C during gestation and abruptly increased by 1–1.5°C on the day of parturition. In free-living females, similar shifts in T _b were observed in 78% (n = 9) of yearlings and 94% (n = 31) of adults; females without the shift are assumed not to have given birth. Three of four ground squirrels for which dates of emergence from hibernation were visually confirmed did not exhibit obvious diurnal rhythms in T _b until they first emerged onto the surface when T _b patterns became diurnal. In free-living males undergoing reproductive maturation, this pre-emergence euthermic interval averaged 20.4 days (n = 56). T _b-loggers represent a cost-effective and logistically feasible method to precisely investigate the phenology of reproduction and hibernation in ground squirrels.     

Summer hibernation induced in ground squirrels (Citellus tridecemlineatus) by urine or plasma from hibernating bats (Myotis lucifugus or Eptesicus fuscus)

Summer hibernation induced in ground squirrels (Citellus tridecemlineatus) by urine or plasma from hibernating bats (Myotis lucifugus or Eptesicus fuscus). Summer hibernation in the thirteen-lined ground squirrel can be induced by intravenous injection of urine or blood plasma previously isolated from winter hibernating little brown bats (M. lucifugus) or big brown bats (E. fuscus). Urine- and plasma-injected ground squirrels kept at 8 °C hibernated earlier, longer, and deeper (as indicated by core temperature and respiratory rate measurements) than control ground squirrels injected with saline. This successful cross-order induction of hibernation demonstrates that the hibernation-inducing trigger (HIT) may be present in nonrodent mammals.     

Seasonal and temperature-related changes in mitochondrial membranes associated with torpor in the mammalian hibernator Spermophilus richardsonii

Seasonal variations in the thermal response of liver mitochondrial membranes from Richardson's ground squirrels (Spermophilus richardsonii) were determined by measuring succinate-cytochrome $\text{c}$ reductase activity and spin label motion over a temperature range of 2 °C to 35 °C. For seven summer animals from the field the Arrhenius-type plots for enzyme activity and spin label motion were biphasic indicating a transition in structure and function at $\text{22 + 2.3°C}\text{and}\text{23 ± 1.9°C}$, respectively; typical of homeothermic mammals. For 12 winter animals maintained at 19°C, the transition in structure and function was lowered to $\text{12 ± 1.1°C}$ and $\text{13 ± 1.4°C}$, respectively. The transition for 5 of 11 winter animals which were kept at 4°C and maintained normal activity and body temperature was similar to animals maintained at 19°C, while for the other six the transition was further lowered to less than 4°C. The transition for seven winter animals which were in deep hibernation was less than 4°C. The results for liver mitochondria show that lowering of the transition in membrane structure and function occurs as a two-stage process of about 10 deg. C for each stage and that the lowering is a requisite for hibernation rather than a response to the low-body temperatures experienced during hibernation.     

Energetics of arousal episodes in hibernating arctic ground squirrels

Arctic ground squirrels overwintering in northern Alaska experience average soil temperature of −10°C. To examine energetic costs of arousing from hibernation under arctic compared to temperate conditions, captive ground squirrels were maintained in ambient temperatures (T _a) of 2, −5 and −12°C. Rates of oxygen consumption and carbon dioxide production were used to estimate metabolic rate and fuel use during the three phases of arousal episodes: rewarming, euthermia, and recooling. Respiratory quotient comparisons suggest exclusive use of lipid during rewarming and mixed fuel use during euthermia. Animals rewarming from torpor at T _a −12°C took longer, consumed more oxygen, and attained higher peak rates of oxygen consumption when compared to 2°C. T _a had no significant effect on cost or duration of the euthermic phase. Animals recooled faster at −12°C than at 2°C, but total oxygen consumption was not different. T _a had no significant effect on the total cost of arousal episodes when all three phases are included. Arousal episodes account for 86% of estimated costs of a complete hibernation cycle including torpor when at 2°C and only 23% at −12°C. Thus, due to the higher costs of steady-state metabolism during torpor, proportional metabolic costs of arousal episodes at T _a characteristic of the Arctic are diminished compared to relative costs of arousals in more temperate conditions.     

Effect of Isoproterenol on the L-type Ca2+ Current in Cardiac Cells from Rats and Hybernating Ground Squirrels

The perforated patch clamp method was used to study the effect of the agonist of beta-adrenoreceptors isoproterenol on L-type Ca²⁺ current in cardiocytes of rats and ground squirrels in two states: active and hibernating. It is shown that isoproterenol exerts a dual effect on Ca²⁺ currents of rats and ground squirrels in the active state: at V _h = –50 mV, the current increases, whereas at V _h = –30 mV, it decreases. In hibernating ground squirrels, the dual effect of isoproterenol is not observed: isoproterenol increases Ca²⁺ current at any V _h values. The hypothesis is put forward that, during the entrance of ground squirrels into hibernation, the phosphorylation of one of the sites (not cAMP-dependent) of L-type Ca²⁺ channels is blocked.     

The optimal depot fat composition for hibernation by golden-mantled ground squirrels (Spermophilus lateralis)

Golden-mantled ground squirrels (Spermophilus lateralis) are herbivores that hibernate during winter. Although little is known about the nutritional/physiological constraints on hibernation, numerous studies have demonstrated that increasing the amount of linoleic acid (a polyunsaturated fatty acid) in the diet enhances hibernation. This is probably because high linoleic acid diets reduce the melting points of the depot fats produced for hibernation which makes them more metabolizable at low body temperatures. This suggests that a major limitation on hibernation may be obtaining enough linoleic acid in the diet for proper hibernation. In all previous studies, however, the amount of linoleic acid in the diets of free-ranging animals was either not considered, or the range of dietary linoleic acid contents in the experiments was less than that of natural diets. It is thus not known whether the amount of linoleic acid available to hibernators under natural conditions actually limits their torpor patterns. A series of laboratory feeding and hibernation experiments were conducted with S. lateralis and artificial diets with different linoleic acid contents that were either below or above the linoleic acid content of the natural diet. The results demonstrated that when dietary linoleic acid contents are either below or above natural levels, hibernation ability is greatly reduced. Hibernation ability was reduced when the squirrels were maintained on a high linoleic acid diet probably by the production of toxic lipid peroxides in brown adipose tissues. The results indicate that there is an optimal level of dietary linoleic acid for proper hibernation, and this is equal to that of the natural diet. The amount of linoleic acid available in the diet thus does not limit hibernation under normal natural conditions.Abbreviations BAT brown adipose tissue - bm body mass - FA fatty acid - PUFA polyunsaturated fatty acid - T _a ambient temperature - T _b body temperature - WAT white adipose tissue     

On the growth of incisors in ground squirrels (Spermophilus) during hibernation

The growth of incisors was studied in two Spermophilus parryii and two S. undulatus ground squirrels with DS-1922L temperature data loggers implanted in the peritoneal cavity, which were kept under laboratory conditions. Daily increments on the incisors surface were similar to those in other species of ground squirrels, but they were less distinct and regular than in wild-living conspecific individuals from the same region. Two S. parryii and one S. undulatus ground squirrels entered hibernation and successfully overwintered. Despite some anomalies in their incisors, changes in body temperature during hibernation (recorded by the data loggers) had an effect on the pattern of their growth, resulting in the formation of a “hibernation zone” on the incisor surface. The number of narrow increments within this zone roughly corresponded to the number of alternating periods of torpor and euthermia during hibernation. This could be regarded as evidence that the incisors of the animals studied continued growing throughout hibernation, including the period of deep hibernation, with the rhythm of their growth coinciding with the rhythm of changes in body temperature. The effect of spontaneous trauma of an upper incisor on the growth of other incisors is described.     

Evaluation of the role of AMP-activated protein kinase and its downstream targets in mammalian hibernation

Mammalian hibernation requires an extensive reorganization of metabolism that typically includes a greater than 95% reduction in metabolic rate, selective inhibition of many ATP-consuming metabolic activities and a change in fuel use to a primary dependence on the oxidation of lipid reserves. We investigated whether the AMP-activated protein kinase (AMPK) could play a regulatory role in this reorganization. AMPK activity and the phosphorylation state of multiple downstream targets were assessed in five organs of thirteen-lined ground squirrels (Spermophilus tridecemlineatus) comparing euthermic animals with squirrels in deep torpor. AMPK activity was increased 3-fold in white adipose tissue from hibernating ground squirrels compared with euthermic controls, but activation was not seen in liver, skeletal muscle, brown adipose tissue or brain. Immunoblotting with phospho-specific antibodies revealed an increase in phosphorylation of eukaryotic elongation factor-2 at the inactivating Thr56 site in white adipose tissue, liver and brain of hibernators, but not in other tissues. Acetyl-CoA carboxylase phosphorylation at the inactivating Ser79 site was markedly increased in brown adipose tissue from hibernators, but no change was seen in white adipose tissue. No change was seen in the level of phosphorylation of the Ser565 AMPK site of hormone-sensitive lipase in adipose tissues of hibernating animals. In conclusion, AMPK does not appear to participate in the metabolic re-organization and/or the metabolic rate depression that occurs during ground squirrel hibernation.     

Diets of native and introduced tree squirrels in Washington

Competition for food resources can be a primary mechanism for displacement of native species by introduced species. Investigation of dietary partitioning between potential competitors and spatiotemporal variability in food resources can reveal contested food items and intensity of competition. Introduced eastern gray squirrels (Sciurus carolinensis) have been implicated as competitors with western gray squirrels (S. griseus) and Douglas’ squirrels (Tamiasciurus douglasii), but little is known about interactions among these species. We collected foraging observations and fecal pellets of sympatric gray and Douglas’ squirrels trapped and radio-tracked in western Washington, USA, from 2007 to 2012. We compared food resources consumed across species, seasons, and years to evaluate dietary overlap and the potential for competitive interactions. All squirrel species ate hypogeous fungi throughout all seasons; spores of several genera were present in 272 of 275 fecal samples and observed in all months. Rhizopogon, Geopora, and Melanogaster spp. occurred in most fecal pellets for all squirrels, but the diet of the Douglas’ squirrels had slightly lower richness of hypogeous fungi than gray squirrels. Although foraging observations suggest that eastern and western gray squirrels shared important dietary items such as acorns, strong differences in consumption of seeds of a common conifer may facilitate their coexistence. Our results suggest that dietary overlap among arboreal squirrels could lead to competitive interactions during periods of food scarcity, but subtle differences may be sufficient to permit long-term coexistence. Tree squirrels in Washington would likely benefit from forest management practices that promote or sustain robust crops of hypogeous fungi. © 2019 The Wildlife Society.     

Hibernation enhancesd-glucose uptake by intestinal brush border membrane vesicles in ground squirrels

The ability to actively transport nutrients is maintained in intestinal tissues of hibernating ground squirrels compared with their active counterparts, and shows apparent upregulation in hibernators when transport rates are normalized to tissue mass. To identify the mechanisms responsible for the preservation of transport function during the extended fast of hibernation, we studiedd-glucose uptake into jejunal brush border membrane vesicles prepared from active and hibernating 13-lined ground squirrels. Hibernators were without food and showing regular bouts of torpor for at least 6 weeks before sacrifice. Electron micrographs indicated similar microvillus heights of jejunal enterocytes in the two activity states, whereas microvillus density was slightly greater in the hibernators. Glucose uptake into brush border membrane vesicles was inversely related to medium osmolarity, indicating negligible binding of substrate to brush border membrane vesicles surfaces, and intravesicular spaces were similar in hibernating and active squirrels. Glucose uptake showed strong Na⁺ dependency in both groups, with equivalent overshoot values in the presence of Na⁺. Kinetic analysis revealed a significant increase in the maximal velocity of transport (J _max) in hibernators (55.9±5.6 nmol·min^-1·mg^-1) compared with active squirrels (36.7±5.1 nmol·min^-1·mg^-1,P<0.05), with no change inK _m. Thus, the structure and absorptive capacity of the intestinal brush border persists in fasted hibernators, and the increase inJ _max for glucose uptake during hibernation likely contributes to the enhanced Na⁺-dependent glucose absorption previously observed at the tissue level.Abbreviations BBM brush border membrane(s) - BBMV brush border membranes vesicles - SGLT1 Na⁺-glucose transporter - 3-OMG 3-orthomethylglucose - J _max maximal velocity of transport - K _m transporter affinity for substrate - T _b body temperature     

Absence of evidence for a hibernation "trigger" in blood dialyzate of Richardson's ground squirrel.

In order to examine evidence for a blood-borne “trigger” for mammalian hibernation, serum dialyzate from hibernating Richardson's ground squirrels (Spermophilus richardsonii) was injected into summer-active ground squirrels of the same species. Four independent trials involving 52 animals were performed. In all trials, no effect of the dialyzate was seen on nest building, weight gain or loss, or on occurrence of hibernation.