Electrophysiological characteristics of cardiac function and the intensity of protein synthesis in cardiomyocytes during the arousal of susliks from hibernation

It has been demonstrated that during winter hibernation (body temperature 2-4 degrees C), the heart rate in ground squirrels is equal to 100 10-12 beats/min. At the initial stage of the arousal, while body temperature remains still low (9-10 degrees C), the heart rate may increase up to 160-200 beats/min. At this stage, practically all electrophysiological parameters of the heart correspond to those in active animals. These results may indicate the ability of "cold" heart in arousing ground squirrels to operate as a normothermic organ and reveal certain role of the heart in body warming. Significant increase of the intensity of protein synthesis in cardiomyocytes together with periodic changes in protein composition of their membranes were found during arousal which may account for regulation of the level of metabolism in cells and for adaptation of the latter to different temperatures.     

The Dynamics of Adaptive Changes in the Spleen of the Hibernating Ground Squirrel <Emphasis Type="Italic">Spermophilus undulatus</Emphasis>

In hibernation season during torpor bouts, the spleen weight and the hemoglobin level, as well as the total and extracted protein contents in the spleen of the ground squirrel Spermophilus undulatus are increased when animals enter torpor and reach maximum values when the body temperature drops below 25°C. All these parameters return to the characteristic values of the euthermic animals during arousal, before the body temperature increases to 20°C. There were no significant differences in the numbers of splenocytes between ground squirrels in interbout euthermia and torpor. The minimum number of splenocytes was observed in animals that entered torpor when the core body temperature was approximately 18°C. The activity of ornithine decarboxylase, a key enzyme in polyamine synthesis, which is correlated with the functional and proliferative status of lymphoid tissue, was the same for the euthermic and summer ground squirrels and decreased monotonically during torpor. Upon arousal of the animals when body temperature was below 29°C, no resumption of the spleen ornithine decarboxylase activity was observed.     

Temperatures in Pigs During 3 T MRI Temperatures,Heart Rates,and Breathing Rates of Pigs During RF Power Deposition in a 3 T (128 MHz) Body Coil

Exposure to radiofrequency (RF) power deposition during magnetic resonance imaging (MRI) induces elevated body‐tissue temperatures and may cause changes in heart and breathing rates, disturbing thermoregulation. Eleven temperature sensors were placed in muscle tissue and one sensor in the rectum (measured in 10 cm depth) of 20 free‐breathing anesthetized pigs to verify temperature curves during RF exposure. Tissue temperatures and heart and breathing rates were measured before, during, and after RF exposure. Pigs were placed into a 60‐cm diameter whole‐body resonator of a 3 T MRI system. Nineteen anesthetized pigs were divided into four RF exposure groups: sham (0 W/kg), low‐exposure (2.7 W/kg, mean exposure time 56 min), moderate‐exposure (4.8 W/kg, mean exposure time 31 min), and high‐exposure (4.4 W/kg, mean exposure time 61 min). One pig was exposed to a whole‐body specific absorption rate (wbSAR) of 11.4 W/kg (extreme‐exposure). Hotspot temperatures, measured by sensor 2, increased by mean 5.0 ± 0.9°C, min 3.9; max 6.3 (low), 7.0 ± 2.3°C, min 4.6; max 9.9 (moderate), and 9.2 ± 4.4°C, min 6.1, max 17.9 (high) compared with 0.3 ± 0.3°C in the sham‐exposure group (min 0.1, max 0.6). Four time‐temperature curves were identified: sinusoidal, parabolic, plateau, and linear. These curve shapes did not correlate with RF intensity, rectal temperature, breathing rate, or heart rate. In all pigs, rectal temperatures increased (2.1 ± 0.9°C) during and even after RF exposure, while hotspot temperatures decreased after exposure. When rectal temperature increased by 1°C, hotspot temperature increased up to 42.8°C within 37 min (low‐exposure) or up to 43.8°C within 24 min (high‐exposure). Global wbSAR did not correlate with maximum hotspot. Bioelectromagnetics. 2021;42:37–50. © 2020 The Authors. Bioelectromagnetics published by Wiley Periodicals LLC on behalf of Bioelectromagnetics Society     

The relative roles of the parasol-like tail and burrow shuttling in thermoregulation of free-ranging Cape ground squirrels,Xerus inauris

As small arid-zone mammals, Cape ground squirrels (Xerus inauris) are unusual in being diurnally active. It is postulated that they remain active during the day by using their parasol-like tails to shade their bodies whilst foraging. However, no studies have continuously measured body temperature to determine the effect of using the tail as a parasol, relative to other thermoregulatory behaviours, such as burrow retreat. We caught four free-ranging Cape ground squirrels (673 ± 36 g) and surgically implanted miniature temperature-sensitive data loggers into their abdomens, to record body temperature every 5 min to an accuracy of 0.04 °C, before they were released back into their home range and observed for two weeks. Mean daily peak black globe temperature was 41 °C, and daily peak body temperature reached 40 °C. Ground squirrels raised their tails significantly more often at globe temperatures above 30 °C, but raising the tail did not decrease body temperature, nor prevent body temperature rising. Ground squirrels retreated to burrows, at 18 °C, significantly more often at high body temperatures and body temperature dropped 1–2 °C before re-emergence. We believe that the tail was raised to provide thermal comfort during high solar radiation exposure, and that burrow retreat was employed to dissipate a heat load and remain active diurnally.     

Comparison of surface temperature in 13-lined ground squirrel (Spermophilus tridecimlineatus) and yellow-bellied marmot (Marmota flaviventris) during arousal from hibernation

Surface temperatures (Ts) of eight 13-lined ground squirrels and seven yellow-bellied marmots were measured during arousal from hibernation using infrared thermography (IRT) and recorded on videotape. Animals aroused normally in 5 degrees C cold rooms. Body temperatures were recorded during arousal using both cheek pouch and interscapular temperature probes. Warming rate in arousal was exponential. Mean mass specific warming rates show the squirrels warm faster (69.76 degrees C/h/kg) than the marmots (4.49 degrees C/h/kg). Surface temperatures (Ts) for 11 regions were measured every few minutes during arousal. The smaller ground squirrel shows the ability to perfuse distal regions without compromising rise in deep body temperature (Tb). All squirrel Ts's remained low as Tb rose to 18 degrees C, at which point, eyes opened, squirrels became more active and all Ts's rose parallel to Tb. Marmot Ts remained low as Tb rose initially. Each marmot showed a plateau phase where Tb remained constant (mean Tb 20.3+/-1.0 degrees C, duration 9.4+/-4.1 min) during which time all Ts's rose, and then remained relatively constant as Tb again began to rise. An anterior to posterior Ts gradient was evident in the ground squirrel, both body and feet. This gradient was only evident in the feet of the marmots.     

Human thermoregulatory responses during prolonged walking in water at 25, 30 and 35°C

Eight healthy and physically well-trained male students exercised on a treadmill for 60 min while being immersed in water to the middle of the chest in a laboratory flowmill. The water velocity was adjusted so that the intensity of exercise correspond to 50% maximal oxygen uptake of each subject, and experiments were performed once at each of three water temperatures: 25, 30, 35°C, following a 30-min control period in air at 25°C, and on a treadmill in air at an ambient temperature of 25°C. Thermal states during rest and exercise were determined by measuring rectal and skin temperatures at various points, and mean skin temperatures were calculated. The intensity of exercise was monitored by measuring oxygen consumption, and heart rate was monitored as an indicator for cardiovascular function. At each water temperature, identical oxygen consumption levels were attained during exercise, indicating that no extra heat was produced by shivering at the lowest water temperature. The slight rise in rectal temperature during exercise was not influenced by the water temperature. The temperatures of skin exposed to air rose slightly during exercise at 25°C and 30°C water temperature and markedly at 35°C. The loss of body mass increased with water temperature indicating that both skin blood flow and sweating during exercise increased with the rise in water temperature. The rise in body temperature provided the thermoregulatory drive for the loss of the heat generated during exercise. Heart rate increased most during exercise in water at 35°C, most likely due to enhanced requirements for skin blood flow. Although such requirements were certainly smallest at 25°C water temperature, heart rate at this temperature was slightly higher than at 30°C suggesting reflex activation of sympathetic control by cold signals from the skin. There was a significantly greater increase in mean skin and rectal temperatures in subjects exercising on the treadmill in air, compared to those exercising in water at 25°C. Accepted: 22 May 1998     

The hibernating 13-lined ground squirrel as a model organism for potential cold storage of platelets

Hibernating mammals have developed many physiological adaptations to extreme environments. During hibernation, 13-lined ground squirrels (Ictidomys tridecemlineatus) must suppress hemostasis to survive prolonged body temperatures of 4-8°C and 3-5 heartbeats per minute without forming lethal clots. Upon arousal in the spring, these ground squirrels must be able to quickly restore normal clotting activity to avoid bleeding. Here we show that ground squirrel platelets stored in vivo at 4-8°C were released back into the blood within 2 h of arousal in the spring with a body temperature of 37°C but were not rapidly cleared from circulation. These released platelets were capable of forming stable clots and remained in circulation for at least 2 days before newly synthesized platelets were detected. Transfusion of autologous platelets stored at 4°C or 37°C showed the same clearance rates in ground squirrels, whereas rat platelets stored in the cold had a 140-fold increase in clearance rate. Our results demonstrate that ground squirrel platelets appear to be resistant to the platelet cold storage lesions observed in other mammals, allowing prolonged storage in cold stasis and preventing rapid clearance upon spring arousal. Elucidating these adaptations could lead to the development of methods to store human platelets in the cold, extending their shelf life.     

The dependence of the rate of heart contractions in the long-tailed Yakutsk suslik Citellus undulatus on environmental temperature]

Decrease of ambient temperature (Ta) leads to the increase of the heart rate (HR) in active ground squirrels C. undulatus by 5.3/min/1 degree C in summer and by 3.8/min/1 degree C in winter. In a hibernation state, the dependence of the HR on Ta was in a good agreement with equation HT = 2.53.exp.(0.1.Ta). On entering into hibernation and on arousal, the HR change outruns the corresponding body temperature (Tb) change by 1.5-2 hours. A maximum HR level (up to 400/min and more) was registered on arousal when Tb reached 17-20 degrees C. A minimal HR level (4-5/min) was observed during hibernation at Ta 2-5 degrees C. The maximum Ta, at witch C. undulatus was hibernating, reached 23-24 degrees C, the HR being 23-25/min.     

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.     

Temporal and temperature effects on the maximum rate of rewarming from hibernation

During hibernation animals oscillate from near ambient (T(a)) to euthermic body temperatures (T(b)). As animals arouse, the rate of rewarming (RRW) might be expected to simply increase as a function of time. We monitored the T(b) of golden-mantled ground squirrels (Spermophilus lateralis) housed at 4, 8, 12, and 16 degrees C during natural arousals. The maximum RRW, the time required to reach a maximum RRW, and the relative time index all demonstrated negative relationships with T(a). The T(b) corresponding to maximal RRW demonstrated a positive relationship with T(a). Squirrels reached maximal RRW when they had generated 30 to 40% of the heat required to reach a euthermic T(b). These data suggest that arousal is more constrained than expected and that both time and temperature influence the RRW.     

Human Core Temperature Responses during Exercise and Subsequent Recovery: An Important Interaction between Diurnal Variation and Measurement Site

Chronobiological investigations into core temperature during and after exercise can involve ambulatory measurements of intestinal temperature during actual competitions, esophageal temperature measurements in laboratory simulations, or rectal temperature, which can be measured in both the field and laboratory. These sites have yet to be compared during both morning and afternoon exercise and subsequent recovery. At 08∶00 and 17∶00 h, seven recreationally active males exercised at 70% peak oxygen uptake for 30 min and then recovered passively for 30 min. During the experiment, esophageal, rectal, intestinal, and skin temperatures, plus sweat loss, heart rate, and ratings of perceived exertion (RPE), were monitored. We found that the diurnal variation in intestinal temperature responses (0.45±0.32°C; mean±SD) was significantly larger compared with rectal (0.33±0.24°C) and, particularly, esophageal temperature responses (0.21±0.20°C; p= 0.019). This reflected a greater difference of 0.25–0.40°C between the esophagus and the other two sites in the afternoon, compared to inter‐site differences of only 0.13–0.16°C in the morning. Diurnal variation was small for skin temperature, heart rate, sweat loss, and RPE responses during exercise (p>0.05). Our data suggest that the relative differences between intestinal, rectal, and esophageal temperature during exercise and subsequent recovery depend on time of day to the extent that inferences from studies on experimental and applied chronobiology will be affected.     

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.     

Cytoskeletal Regulation Dominates Temperature-Sensitive Proteomic Changes of Hibernation in Forebrain of 13-Lined Ground Squirrels

13-lined ground squirrels, Ictidomys tridecemlineatus, are obligate hibernators that transition annually between summer homeothermy and winter heterothermy – wherein they exploit episodic torpor bouts. Despite cerebral ischemia during torpor and rapid reperfusion during arousal, hibernator brains resist damage and the animals emerge neurologically intact each spring. We hypothesized that protein changes in the brain underlie winter neuroprotection. To identify candidate proteins, we applied a sensitive 2D gel electrophoresis method to quantify protein differences among forebrain extracts prepared from ground squirrels in two summer, four winter and fall transition states. Proteins that differed among groups were identified using LC-MS/MS. Only 84 protein spots varied significantly among the defined states of hibernation. Protein changes in the forebrain proteome fell largely into two reciprocal patterns with a strong body temperature dependence. The importance of body temperature was tested in animals from the fall; these fall animals use torpor sporadically with body temperatures mirroring ambient temperatures between 4 and 21°C as they navigate the transition between summer homeothermy and winter heterothermy. Unlike cold-torpid fall ground squirrels, warm-torpid individuals strongly resembled the homeotherms, indicating that the changes observed in torpid hibernators are defined by body temperature, not torpor per se. Metabolic enzymes were largely unchanged despite varied metabolic activity across annual and torpor-arousal cycles. Instead, the majority of the observed changes were cytoskeletal proteins and their regulators. While cytoskeletal structural proteins tended to differ seasonally, i.e., between summer homeothermy and winter heterothermy, their regulatory proteins were more strongly affected by body temperature. Changes in the abundance of various isoforms of the microtubule assembly and disassembly regulatory proteins dihydropyrimidinase-related protein and stathmin suggested mechanisms for rapid cytoskeletal reorganization on return to euthermy during torpor-arousal cycles.     

Cold tolerance in rats following administration of streptozotocin,glucose, insulin and glucose plus insulin

Fall in rectal temperature (T_re) and survival time was determined on exposure to–20°C in adult normoglycemic and diabetic (streptozotocin treated) rats and 1 h following glucose feeding or insulin administration or both and on exposure to–10°C in young rats with and without glucose feeding. The susceptibility to frostbite was determined by exposure of the limbs to freezing mixture of–19°C or–23°C. The rate of fall of T_re was less and the survival time more in glucose and insulin plus glucose treated animals. On the other hand, the rate of fall of T_re was more and the survival time less, in dia betic and insulin-treated animals. The rectal temperature at which the animal died was the same in the control and the treated animals. The susceptibility to frost bite was more in insulin treated and diabetic animals and less in glucose-fed animals. Exposure to cold during the second h after glucose or glucose plus insulin injection did not alter the blood glucose from that obtained at room temperature. In insulin-treated animals the rate of rise of blood glucose during the second h was much higher at low temperature than at room temperature. The rise in blood glucose in diabetic animals was much higher than in normoglycemic animals exposed to cold.     

Adaptation to low body temperature influences pulmonary surfactant composition thereby increasing fluidity while maintaining appropriately ordered membrane structure and surface activity

The interfacial surface tension of the lung is regulated by phospholipid-rich pulmonary surfactant films. Small changes in temperature affect surfactant structure and function in vitro. We compared the compositional, thermodynamic and functional properties of surfactant from hibernating and summer-active 13-lined ground squirrels (Ictidomys tridecemlineatus) with porcine surfactant to understand structure-function relationships in surfactant membranes and films. Hibernating squirrels had more surfactant large aggregates with more fluid monounsaturated molecular species than summer-active animals. The latter had more unsaturated species than porcine surfactant. Cold-adapted surfactant membranes displayed gel-to-fluid transitions at lower phase transition temperatures with reduced enthalpy. Both hibernating and summer-active squirrel surfactants exhibited lower enthalpy than porcine surfactant. LAURDAN fluorescence and DPH anisotropy revealed that surfactant bilayers from both groups of squirrels possessed similar ordered phase characteristics at low temperatures. While ground squirrel surfactants functioned well during dynamic cycling at 3, 25, and 37°C, porcine surfactant demonstrated poorer activity at 3°C but was superior at 37°C. Consequently the surfactant composition of ground squirrels confers a greater thermal flexibility relative to homeothermic mammals, while retaining tight lipid packing at low body temperatures. This may represent the most critical feature contributing to sustained stability of the respiratory interface at low lung volumes. Thus, while less effective than porcine surfactant at 37°C, summer-active surfactant functions adequately at both 37°C and 3°C allowing these animals to enter hibernation. Here further compositional alterations occur which improve function at low temperatures by maintaining adequate stability at low lung volumes and when temperature increases during arousal from hibernation.     

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.     

Analysis of changes in heart rate and temperature of the ground squirrel Citellus undulatus in various physiological states

Heart rate (HR) of ground squirrel C. undulatus was studied in dependence of season, level of activity, physiological state and air temperature (T). In summer HR varies from 110-130 beat/min in sleep up to 420 beat/min at flight from danger. During winter hibernation HP was minimal (3-5 beat/min) at T 1-4 degrees C, the increase in T induced the growth of HP in correspondence to the Arrenius van't Hoff law. The temperature of the body in hibernation exceeded T on 1.5-3 degrees C. The time of getting off the hibernation increased with the decrease in T (6-7 hours at -1 degree C and 2.5-3 hours at +18 degrees C). At phase of increased thermogenesis during arousal heart temperature exceeded rectal one on 10-12 degrees C and heart rate run up to 360-420 beat/min i.e. 2-3 time higher than in active state. The decrease in T stimulated the increase in HP up to 3.8 in winter and 5.3 beat/min/degree C in summer. The highest values of Q10 for HP were revealed at the beginning of hibernation (15-20) and at the beginning of arousal (6-7), in other periods Q10 was similar to the normal biological values (2-2.5). Thus, at the beginning of transitional periods changes of HP were determined mainly by endogenic mechanisms that inhibited myocardium at the beginning of hibernation and activated in arousal. Some mechanisms of coordination between activities of heart and other systems of organism are considered. The features of hit exchange providing the hibernation in wide range of T are discussed.     

The effect of serotonin on the heart contraction frequency in the suslik Citellus undulatus depends on the environmental temperature

In experiments on active ground squirrels, it has been shown that the effect of intraperitoneal injection of serotonin on the heart rate depends on the dose and the environmental temperature. At relatively low doses and high environmental temperature, serotonin increases the heart rate and body temperature; on the contrary, with the increase in the dosage and the decrease in the environmental temperature the heart rate and body temperature of animals decrease. It is suggested that this relationship serves as one of the factors which enhance the transition of animals from one level of activity to another during the beginning and the end of hibernation.     

Studies on the estrus cycle of the ground squirrel (Citellus citellus L.) at experimentally induced hibernation

Eighty-five ground squirrels (C. citellus L.) were examined immediately after arousal (March-June). It was established that the animals were in estrus at the end of March; in April they were pregnant; in May they were in lactation; and in June they were already at rest, a postlactation period. Fifteen ground squirrels were treated with Hibernal and then placed in a chamber for 30 days at external temperature, 8-12 degrees C. In this case the estrus cycle of hypothermic ground squirrels was not changed. A retarded involution of corpus luteum persisted for a month and cystic follicles appeared in the ovary.