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Early postdenervation depolarization develops faster at endplates of hibernating golden hamsters where spontaneous quantal and non-quantal acetylcholine release is very small.

The hyperpolarization produced by the application of curare to the postsynaptic membrane of the diaphragm neuromuscular synapse (H-effect) is a measure of non-quantal release (NQR) of acetylcholine (ACh) from the motor nerve ending. In mouse diaphragm, H-effect was 9.3 mV, significantly lower in awake hamsters (7.1 mV) and very small (1.1 mV) in hibernating hamsters. Also, the initial resting membrane potential (RMP) after dissection was highest in mouse (81.5 mV, inside negative), significantly smaller in awake hamsters (77.9 mV) and lowest in hibernating hamsters (75.1 mV). The early postdenervation depolarization of muscle fiber RMP to about 66-68 mV developed with half-decay time (T1/2) of 120 min in mouse, more rapidly in active hamsters (T1/2=60 min) and even faster in hibernating hamsters (T1/2=25 min) muscles. This reciprocal correlation between the H-effect and the rate of early depolarization indicates that non-quantal release is important for maintaining the resting membrane potential [Vyskocil, F. 2003. Early postdenervation depolarization is controlled by acetylcholine and glutamate via nitric oxide regulation of the chloride transporter. Neurochem. Res. 28, 575-585]. The amplitude of H-effect in mouse and hamster was proportional to the spontaneous quantal release. The frequency of miniature endplate potentials was highest in mouse (1.6 s-1), much smaller in awake hamsters (0.51 s-1) and very small in hibernating hamsters (0.08 s-1). This is in accordance with the idea that non-quantal release depends on the number of vesicles fused with the presynaptic membrane during quantal release [Edwards et al., 1985; Ferguson, S.M., Savchenko, V., Apparsundaram, S., Zwick, M., Wright J., Heilman, C.J., Yi, H., Levey, A.I., Blakely R.D. Vesicular localization and activity-dependent trafficking of presynaptic choline transporters. J. Neurosci. 23 (2003) 9697-9709].

Acetylcholine↗

Purification and characterization of fructose bisphosphate aldolase from the ground squirrel, Spermophilus lateralis: enzyme role in mammalian hibernation.

Fructose-1,6-bisphosphate (F1,6P(2)) aldolase was purified to homogeneity from skeletal muscle of the golden-mantled ground squirrel, Spermophilus lateralis. Enzyme properties were examined at temperatures characteristic of euthermia (37 degrees C) and hibernation (5 degrees C); parallel studies assessed rabbit muscle aldolase for comparison. Kinetic properties of each enzyme were differentially affected by assay temperature. For example, the K(m) for F1,6P(2) of ground squirrel aldolase was 0.9+/-0.05 microM at 37 degrees C and 50% higher (1.45+/-0.04 microM) at 5 degrees C, whereas the K(m) of rabbit aldolase increased threefold over the same temperature range. The inhibitory effects of adenylates were similar at both temperatures for the ground squirrel enzyme, but inhibition by adenosine 5(')-diphosphate, adenosine 5(')-monophosphate, and inosine 5(')-monophosphate was substantially reduced at 5 degrees C for rabbit aldolase. Inhibition by inorganic phosphate increased at lower temperatures for both enzymes; for ground squirrel aldolase, the K(i) was 1.18+/-0.1mM at 37 degrees C and 0.23+/-0.05 mM at 5 degrees C. Inhibition of aldolase by inorganic phosphate could be one factor that helps to shut down glycolysis during hibernation. Thus, mammalian hibernators may exploit low-temperature characteristics of aldolase to benefit the metabolic needs of the hibernating state.

Adenosine Diphosphate↗

Intrahippocampal histamine delays arousal from hibernation.

Hibernation is a state of extremely reduced physiological functions and a deep depression of CNS activity, which is thought to be under hippocampal control. Our previous findings indicate increased histamine turnover during hibernation in several brain regions, including the hippocampus. In this study we showed that histamine infused into the hippocampus significantly delayed arousal from hibernation. These findings indicate that histamine may contribute to maintaining the hibernating state, suggesting a novel role for histamine in controlling arousal state.

Animals↗

Hibernation-induction trigger. I. Opioid-like effects of prairie dog plasma albumin on induced contractility of guinea pig ileum.

Studies have shown that plasma albumin fractions (PAFs) from hibernating mammals can inhibit induced contractility of the guinea pig ileum similarly to morphine. This study examined PAFs from two species of prairie dogs, one that undergoes natural seasonal hibernation (white-tailed, WT) and one that does not but can be induced to hibernate (black-tailed, BT). Dose-response curves of lyophilized PAF yielded IC50 values (mg) of 20.23 for summer WT, 15.53 for hibernating WT, 15.45 for summer BT, and 13.16 for winter-active BT. Winter samples from both species have IC50s lower than samples from summer animals, indicating greater potency of winter PAFs in suppressing guinea pig ileum contractility and therefore the presence of more opioid ligands in winter prairie dog plasma. Studies to elucidate receptor selectivity of PAF continue.

Animals↗

Effect of hibernation, thyroid hormones and dexamethasone on cytosolic and mitochondrial glycerol-3-phosphate dehydrogenase from jerboa (Jaculus orientalis).

Tissue distribution of the cytosolic and mitochondrial glycerol-3-phosphate dehydrogenase (cGPDH and mGPDH) activities in jerboa (Jaculus orientalis), a hibernator, shows the highest level of enzyme activity in skeletal muscle and brown adipose tissue, respectively. The effect of hibernation on cGPDH indicates an increase of activity in all tissues examined. In contrast, hibernation decreases mGPDH activity in all tissues, except skeletal muscle. The effect of thyroid hormones on GPDH activity was tissue specific: in kidneys, cGPDH activity doubled in euthermic jerboas treated with T4. In contrast, 6-n-propyl-2-thiouracil treatment provokes an increase of enzyme activity in brown adipose tissue, liver and brain. T4 treatment leads to a 2.7-fold increase in liver mGPDH activity. 6-n-propyl-2-thiouracil treatment decreases mGPDH activity in the skeletal muscle whereas the opposite effect was observed in brain. Dexamethasone stimulates cGPDH in all tissues examined, except skeletal muscle and kidneys. In the case of mGPDH activity, this increase was observed only for brown adipose tissue and brain. Our results suggest that hibernation, thyroid hormones and dexamethasone probably play a role in the regulation of cGPDH and mGPDH activities in jerboa. Our findings confirm that these enzymes are involved in metabolic adaptation to thermal stress in Jaculus orientalis.

Adipose Tissue, Brown↗

Liver and kidney structure and iron content in romanian brown bears (Ursus arctos) before and after hibernation.

The annual cycle of the brown bear (Ursus arctos) in the Carpathian Mountains (Romania) consists of an active period from April to November, and an inactive period (hibernation) of approximately 4-5 months between November and March. During hibernation, the brown bears sleep continually and do not feed or drink water. Analyses of liver and kidney of male brown bears showed that liver iron content was 3 times higher in bears at the end of hibernation than at the end of the active period. A possible trend towards a decrease in iron content was noted for the kidney. The presence of iron in the liver was confirmed by the presence of the Perls-positive granules in the cytoplasm of Kupffer cells, in other non-parenchymal cells and also in some hepatocytes. The hepatic veins of the bear liver samples obtained in early spring showed narrower lumens with pleated walls, compared to the normal outline of the hepatic veins in the liver from the bears sampled during autumn. Also in the early spring bears, the renal glomeruli were partially fibrosed. Renal glomerular fibrosis was sometimes observed in samples from the prehibernation period. The tissue iron values from the livers and kidneys of brown bears in early spring or autumn might provide useful data on iron metabolism under conditions of hibernation and accompanying starvation.

Animals↗

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.

Animals↗

Noradrenaline-induced lipolysis in adipose tissue is suppressed at hibernation temperatures in ground squirrels.

Responsiveness of white adipose tissue (WAT) and brown adipose tissue (BAT) from hibernating and nonhibernating golden-mantled ground squirrels (Spermophilus lateralis) to the lipolytic action of the sympathetic neurotransmitter, noradrenaline, was tested in vitro at temperatures characteristic of deep torpor (5 degrees C) and euthermia (37 degrees C). Noradrenaline-stimulated lipolysis, as indicated by WAT glycerol release, of tissue from hibernating ground squirrels was six- to ten-fold greater at 37 degrees C than at 5 degrees C. Noradrenaline was ineffective in increasing lipolysis at 5 degrees C. Noradrenaline-stimulated lipolysis in BAT was similarly suppressed at 5 degrees C. Noradrenaline-stimulated lipolysis was little affected by temperature change below approximately 15 degrees C but strongly correlated with temperature above approximately 15 degrees C. Noradrenaline-induced lipolysis of WAT from nonhibernating and hibernating ground squirrels did not differ at an incubation temperature of 5 degrees C. We conclude that noradrenaline-stimulated WAT lipolytic activity is markedly suppressed at the low temperatures characteristic of deep torpor and that there is no 'hibernation-specific' adaptation of WAT to enhance its responsiveness to noradrenaline at low tissue temperatures. Temperature dependence of noradrenaline-stimulated lipolysis may in part account for the shift from lipid to carbohydrate metabolism during the earliest stage of arousal from deep torpor.

Adipose Tissue↗

Medical significance of cardiovascular function in hibernating mammals.

1. The hibernating mammals inherited a stable cardiovascular function as a result of adaptation to extreme external and internal environments during hibernation. 2. The cardiovascular function of a hibernator is characterized by three distinct features: (i) tolerance of hypothermia; (ii) resistance to dysrhythmia; and (iii) endurance of hypoxia. All these have clinical relevance in cardiovascular disorders in humans. 3. We suggest that the hibernating mammal can serve as a useful animal model for research into the cardiovascular system.

Animals↗

Central vasopressin infusion prevents hibernation in the European hamster (Cricetus cricetus).

The amount of immunocytochemically detectable vasopressin in the brain of the European hamster (Cricetus cricetus) shows a seasonal variation; i.e., dense vasopressin immunoreactivity is present in the lateral septum during summer but is absent in autumn and winter [Buijs, R. M., Pévet, P., Masson-Pévet, M., Pool, C. W., De Vries, G. J., Canguilhem, B. & Vivien-Roels, B. (1986) Brain Res. 371, 193-196]. In the winter period the European hamster hibernates. Since vasopressin in the lateral septum is known to be involved in the control of body temperature, we investigated whether infusion of vasopressin in the lateral septum during autumn-winter could influence hypothermic patterns normally seen in hibernating animals. Hamsters whose lateral septum was infused with vasopressin showed almost no periods of hypothermia, whereas hamsters treated with control infusions displayed a normal hibernation pattern. The results indicate that persistence of vasopressin release in the lateral septum of the European hamster during winter can prevent hibernation.

Animals↗

Effects of changing ambient temperature on metabolic, heart, and ventilation rates during steady state hibernation in golden-mantled ground squirrels (Spermophilus lateralis).

To determine whether metabolic rate is suppressed in a temperature-independent fashion in the golden-mantled ground squirrel during steady state hibernation, we measured body temperature and metabolic rate in ground squirrels during hibernation at different T(a)'s. In addition, we attempted to determine whether heart rate, ventilation rate, and breathing patterns changed as a function of body temperature or metabolic rate. We found that metabolic rate changed with T(a) as it was raised from 5 degrees to 14 degrees C, which supports the theory that different species sustain falls in metabolic rate during hibernation in different ways. Heart rate and breathing pattern also changed with changing T(a), while breathing frequency did not. That the total breathing frequency did not correlate closely with oxygen consumption or body temperature, while the breathing pattern did, raises important questions regarding the mechanisms controlling ventilation during hibernation.

Adaptation, Physiological↗

The role of energy availability in Mammalian hibernation: an experimental test in free-ranging eastern chipmunks.

Reduced torpor expression by hibernating mammals is often attributed to physiological constraints that limit their hibernation ability but may instead reflect adaptive, plastic responses to surplus energy availability. We evaluated this hypothesis by supplementing the food hoards of free-ranging eastern chipmunks (Tamias striatus) before hibernation and then documenting their use of torpor during the subsequent winter. In both years of study, chipmunks that received additional food were euthermic more than twice as frequently as nonsupplemented individuals. Furthermore, when food-supplemented individuals did express torpor, their minimum collar temperature was 5 degrees -10 degrees C warmer than nonsupplemented animals. These results indicate that reduced torpor expression by hibernators can result from an absence of energetic necessity rather than a lack of physiological capability and suggest that even endotherms sequestered in a hibernaculum may benefit from maintaining an elevated body temperature whenever possible.

Animals↗

Mitochondrial metabolism in hibernation: metabolic suppression, temperature effects, and substrate preferences.

We compared liver and skeletal muscle mitochondrial function among activity states to characterize regulated reversible metabolic suppression in the mammalian hibernator Spermophilus tridecemlineatus. At 37 degrees C, succinate oxidation was 70% lower in the liver mitochondria from torpid animals than in those from summer-active animals or in animals arousing from torpor. Respiration was very sensitive to temperature (Q(10) 5.8-9.8), and when measured at 25 degrees or 5 degrees C there was no difference among the three states. Liver mitochondria from summer-active animals oxidized pyruvate and beta -hydroxybutyrate at higher rates than those from torpid animals, and flux through complex 4 of the electron transport chain was about three- and fivefold higher than flux through complexes 2-4 and complexes 1-4, respectively. In the hibernating and arousing animals there was no difference in flux through complexes 2-4 and complex 4, suggesting a downregulation of cytochrome c oxidase in liver mitochondria during the hibernation season. Muscle mitochondrial respiration did not differ between the torpid and summer-active states in any of the parameters measured. The data support a regulated, reversible decrease of liver (but not muscle) mitochondrial oxidative phosphorylation in hibernating ground squirrels.

3-Hydroxybutyric Acid↗

Fat ablation and food restriction influence reproductive development and hibernation in ground squirrels.

The role of white adipose tissue in development of the reproductive apparatus of male golden-mantled ground squirrels was assessed by surgical removal of fat (lipectomy) immediately prior to onset of hibernation or by manipulation of fat levels through food restriction for 5 wk preceding hibernation. Animals then were maintained without food at 6 degrees C, and they hibernated from November 1983 until April 1984. At that time, blood plasma was assayed for hormone levels, and body mass, body composition, and masses of the testes and seminal vesicle-prostate complex were determined. At autopsy, testes and sexual accessory organs were heavier in Control squirrels than in Food-Restricted or Lipectomized (LIPX) animals. Paired-testes mass was positively correlated with body mass. Testosterone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) levels were uniformly low in animals sampled during torpor, but were elevated in 2 animals that had regained the euthermic state. The Food-Restricted animals spent more time in torpor than did the other two groups, and the percentage of decrease in body mass over the hibernation season was less for Food-Restricted than for LIPX or Control animals. Adequate lipid stores may be essential for the normal development of the reproductive apparatus of male ground squirrels. In the field, animals with insufficient fat stores may forego reproduction and thereby conserve energy for survival.

Adipose Tissue↗

Hibernation induced in ground squirrels by blood transfusion.

Natural mammalian hibernation was continuously maintained under laboratory conditions throughout spring and summer seasons in a colony of thirteen-lined ground squirrels by serial transfusional passage of blood from hibernations animals to active animals. This procedure successfully produced hibernation in animals until late summer, at which time naturally occurring (spontaneous) hibernations occurred in the colony, thus terminating the experiment.

Animals↗

Summer hibernation induced by cryogenically preserved blood "trigger".

Summer hibernation induced by transfusions of blood (serum, cells, or whole blood) from hibernating ground squirrels and woodchucks demonstrates that (i) this "trigger effect" can be preserved cryogenically in vitro for at least 5 months, (ii) it cross-reacts between these species of hibernators, and (iii) its effectiveness relates to the donor's previous hibernation history.

Animals↗

Classification and distribution of large intestinal bacteria in nonhibernating and hibernating leopard frogs (Rana pipiens).

The large intestinal flora of the leopard frog, Rana pipiens, was examined to determine whether differences existed between the nonhibernating and hibernating states of the animal and to determine the relative concentrations and proportions of potential frog pathogens. Hibernators had a logarithmic decrease of bacteria per milligram of intestine averaging one, and significantly greater proportions of facultative bacteria and psychrophiles relative to nonhibernators. The predominant anaerobic bacteria were gram-positive Clostridium species and gram-negative Bacteroides and Fusobacterium species. The predominant facultative bacteria were enterobacteria in nonhibernators but Pseudomonas species in hibernators. Many species of Pseudomonas are pathogenic for frogs, and thus the intestinal flora in hibernators may be a potential source of infectious disease.

Anaerobiosis↗

mRNA stability and polysome loss in hibernating Arctic ground squirrels (Spermophilus parryii).

All small mammalian hibernators periodically rewarm from torpor to high, euthermic body temperatures for brief intervals throughout the hibernating season. The functional significance of these arousal episodes is unknown, but one suggestion is that rewarming may be related to replacement of gene products lost during torpor due to degradation of mRNA. To assess the stability of mRNA as a function of the hibernation state, we examined the poly(A) tail lengths of liver mRNA from arctic ground squirrels sacrificed during four hibernation states (early and late during a torpor bout and early and late following arousal from torpor) and from active ground squirrels sacrificed in the summer. Poly(A) tail lengths were not altered during torpor, suggesting either that mRNA is stabilized or that transcription continues during torpor. In mRNA isolated from torpid ground squirrels, we observed a pattern of 12 poly(A) residues at greater densities approximately every 27 nucleotides along the poly(A) tail, which is a pattern consistent with binding of poly(A)-binding protein. The intensity of this pattern was significantly reduced following arousal from torpor and undetectable in mRNA obtained from summer ground squirrels. Analyses of polysome profiles revealed a significant reduction in polyribosomes in torpid animals, indicating that translation is depressed during torpor.

Animals↗