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Endogenous opioid system down-regulation during hibernation in amphibians.

In late summer, Northern grass frogs, Rana pipiens, intraspinally administered dynorphin produces a potent, dose-dependent antinociceptive action as measured by the acetic acid test used to evoke a hindlimb wiping response. Surprisingly, in fall, frogs which have entered hibernation, intraspinal dynorphin produces no antinociception action. Intraspinal morphine shows a decreased effect in fall frogs while systemic morphine is equi-effective in summer and fall frogs. Immobilization stress, previously shown to be mediated by endogenous opioid systems in this amphibian, produces a robust increase in nociceptive thresholds in summer frogs while the nociceptive thresholds of fall frogs are unaffected by this procedure. Summer frogs adapted to cold room (4 degrees C) show a significant decrease in nociceptive thresholds compared to cohorts kept at room temperature, and cold-adapted frogs returned to room temperature show a naloxone-attenuated increase in nociceptive threshold. Collectively, these data suggest that endogenous opioid systems in these northern frogs are down-regulated during fall hibernation.

Animals↗

Olfactory bulb removal lengthens the period of circannual rhythms and disrupts hibernation in golden-mantled ground squirrels.

Removal of the olfactory bulbs lengthened the period of circannual rhythms (CARs) of body mass and plasma testosterone (T) concentrations in male golden-mantled ground squirrels, but did not otherwise alter their expression. The period of the CAR was approximately 2 months longer in bulbectomized than in sham-operated animals. Peak values of body mass and T were unaffected by bulbectomy. All neurologically intact, but only 50% of bulbectomized, squirrels displayed normal hibernation patterns. We conclude that the olfactory bulbs are not the site of essential circannual oscillators. Circannual reproductive and hibernation cycles of ground squirrels are less subject to modulation by the olfactory bulbs than are the corresponding rhythms of several non-circannual hamster species.

Animals↗

Fluorescence anisotropy of kidney lipids and membranes of a hibernating mammal.

The fluorescence anisotropy of lipids and membranes isolated from kidneys of European hamsters (Cricetus cricetus L.) has been estimated using 1,6-diphenyl-1,3,5-hexatriene as a probe. We have compared in this study the results obtained for two critical periods for a hibernator: winter (torpid state), and summer (active state). The differences were of very low magnitude. A slight increase in anisotropy was noticed in the kidney lipids and microsomal membrane preparations from torpid animals. In contrast, a small decrease in anisotropy was observed in the microsomal lipid extracts of torpid animals. A difference in triglyceride content of winter and summer total kidney lipids was detected, as well as a difference in microsomal protein content between winter and summer membrane preparations. It is hypothesized that the latter observations may explain why the behavior of kidney total lipids and microsomal preparations were different from that presented by kidney microsomal lipids in respect to fluorescence anisotropy. Therefore, only a little, if any, homeoviscous adaptation is exhibited by kidney membranes during hibernation of this mammal.

Animals↗

The influence of cholecystokinin octapeptide and angiotensin II on in vitro duodenal motility in hibernating and aroused 13-lined ground squirrels.

Because cholecystokinin octapeptide and angiotensin II are directly involved in intestinal food and water absorption, the effect of these two compounds on intestinal motor responses of hibernating and alert 13-lined ground squirrels was investigated. Both cholecystokinin octapeptide and angiotensin II caused a greater increase in the composite motility of intestinal segments of normothermic in contrast to hypothermic hibernating ground squirrels. Additionally, cholecystokinin caused an increase in the contraction frequency of the intestine from normothermic as compared to hypothermic squirrels. This differential response may provide an adaptive advantage by decreasing food and water consumption; depressing motility and inhibiting digestive enzyme release; reducing the release of bile which also may have an irritant effect on the intestinal mucosa under prolonged exposure. As a result, the GI tract is devoid of food and digestive enzyme irritants during torporous periods.

Angiotensin II↗

Histoenzymatic activity of the delta 5-3 beta-hydroxysteroid dehydrogenase of ovary in hibernators (Citellus citellus L).

The ovaries of 24 ground squirrels (Citellus citellus L) were studied in spring (March, April), summer (July), and winter (December). The animals hibernated in a chamber at a temperature of 6-8 degrees C. The activities of the interstitial gland, theca interna, atretic follicles with theca interna and yellow bodies were measured densitometrically and studied by a specially modified histoenzymatic technique. Measurements showed that the endocrine structures were most active in March and least active in July. The atretic follicles had the highest enzymatic activity in April. The quantitative histoenzymatic approach presents an objective base for morphofunctional studies of organs and tissues at different body temperature levels in hibernators.

3-Hydroxysteroid Dehydrogenases↗

Plasma levels of atrial natriuretic factor in nonhibernating and hibernating marmots.

Plasma atrial natriuretic factor (ANF) was measured in 16 marmots at various times of the year. Nonhibernating males (n = 6) had an average plasma concentration of 56 +/- 8 pg/ml; nonhibernating females (n = 6) had an average plasma concentration of 61 +/- 4 pg/ml. During hibernation an additional group of females (n = 4) showed an average of 25 +/- 5 pg/ml. Plasma ANF of both groups of nonhibernating marmots was significantly higher (P less than 0.01) than that the hibernating group, but there was no difference between nonhibernating males and females.

Animals↗

Peculiarities of functioning of protein-synthesizing apparatus of the hibernator (Citellus undulatus).

Synthesis of proteins and RNA has been studied by means of labeled precursors of 14C-labeled amino acids and [14C]orotic acid in liver cells of hibernating, arousing, and active ground squirrels (Citellus undulatus). The investigations indicate that cycloheximide (CHI)-inhibited protein synthesis in liver cells of hibernating ground squirrels may occur on preexisting polyribosomes. A low level of protein synthesis in this state results not only from the Arrhenius dependence, but also from the dissociation of polyribosomes into monosomes. During arousal there is an intensive assembly of polyribosomes. This occurs even with the inhibition of RNA synthesis by actinomycin D, testifying to the presence of a pool of iRNA in liver cells, which is necessary to activate translation. The intensity of protein synthesis in arousal is also influenced by the activation of transcription.

Animals↗

Cyclosporin A suppression of uncoupling in liver mitochondria of ground squirrel during arousal from hibernation.

Energy coupling parameters were studied in liver mitochondria of ground squirrel during arousal from hibernation. It was found that such mitochondria become uncoupled during incubation with phosphate in a salt medium. The uncoupling was revealed by respiration rate increase and membrane potential decrease in the presence of oligomycin. Both effects were reversed by addition of cyclosporin A. Under the same in vitro conditions, mitochondria from aroused (active) animals showed no uncoupling but could be uncoupled by addition of palmitate in the cyclosporin A-sensitive fashion. It is proposed that formation of cyclosporin A-sensitive pores can be involved in urgent heat production in arousing hibernators.

Animals↗

Annual plasma testosterone and thyroxine cycles in relation to hibernation in the edible dormouse Glis glis.

Twelve male edible dormice, captured in the autumn of 1979, were studied for 2 years under natural lighting in an animal enclosure with permanently open windows, at Montpellier, in Southern France (latitude 43 degrees 50' N). Ambient temperature fluctuated annually between +4 and 27 degrees, and the photoperiod between 10L:14D and 16L:8D. Another group of 26 animals, captured in the autumn of 1980, was observed under the same conditions for 1 year. Body weight and plasma testosterone and thyroxine concentrations were measured at monthly intervals under standardized conditions. Hibernation generally extended from October to February but lasted till May in a few cases, and the critical temperature for dormancy varied between 13 and 16 degrees in November and 23 degrees in May. Body weight fluctuated annually according to a biphasic pattern, with two maxima, respectively, in spring and autumn (prehibernation), and two minima, in summer and winter. However, plasma testosterone and thyroxine concentrations displayed clearly monophasic annual cycles, closely correlated, both to each other and to the ambient temperature and photoperiod. Both the testosterone and thyroxine cycles culminated in June-July. Interestingly, the annual ascending phase for both hormonal cycles never preceded the terminal arousal of the animal. Further, whatever the season, 30 degrees was clearly the critical minimal body temperature required for a seasonal increase to occur in plasma testosterone. These annual cycles are discussed in the light of the data acquired recently for hibernators other than the dormouse.

Animals↗

Hibernation depth influences the edible dormouse pancreatic B cell during the spring arousal.

In order to determine the influence of hibernation depth upon the secretion and the effect of insulin, two groups of edible dormice were maintained in winter under different climatic and nutritional conditions, and their pancreatic B-cell function was tested during the spring arousal. The first group of animals was exposed to a moderate temperature and fed ad libitum. Their periods of hypothermia were short and irregular and the active periods sometimes lasted several days; their body weight increased during the winter months; in spring, the sensitivity of B cells to glucose was low, decreasing insulin secretion in vivo and in vitro, and the adipocytes were insulin resistant. The second group of fasting animals was exposed to a low and constant temperature (5 degrees). Their phases of lethargy were long and regular (about 15 days), separated by active periods (6-8 hr); their body weight decreased during the winter months; in spring the B-cell secretion was increased and the sensitivity of the tissues to insulin ensured a high peripheral glucose utilization. These data show that the winter climatic and nutritional conditions which influence the depth of hibernation modify the edible dormouse B-cell activity during the spring arousal.

Adipose Tissue↗

Morphine physical dependence in the hibernator: central nervous system mechanisms underlying the development of dependence remain functional during depression induced by pentobarbital anesthesia.

Earlier work suggested that adaptive mechanisms of the hibernating brain may also block the development of morphine physical dependence. To test an alternate view that CNS depression itself might be the major factor in the failure of dependence to develop, we compared the strength of dependence produced by morphine (two 75-mg pellets, s.c.) given for 12 hr in the presence, versus in the absence, of continuous pentobarbital anesthesia in nonhibernating ground squirrels (Citellus lateralis) and, in addition, in rats. Dependence was measured by the naloxone (5 mg/kg, s.c.) evoked abstinence syndrome in the awake state. The results demonstrated that pentobarbital-induced general anesthesia does not significantly reduce the development of morphine dependence in either species. We conclude that CNS depression alone does not account for the hibernation-related reduction in morphine physical dependence.

Anesthesia↗

Hibernation "trigger" alters renal function in the primate.

Macaque monkeys acclimatized to a restraint chair were fitted with indwelling venous and urinary catheters. After basal rates of urine production and creatinine clearance were determined, a 50 mg dose of plasma dialysate albumin fraction obtained from the woodchuck was administered intravenously in a total volume of 2.5 ml. Plasma fractions were collected during the winter interval of hibernation (hibernation "trigger" or HT), or during the summer active (SAWA) period. Although the SAWA fraction exerted no effects on renal function, HT caused a significant reduction in creatinine clearance. In addition, a tendency toward reduced urine flow and creatinine production occurred following the HT infusion. These findings suggest that over and above the hypothermia, aphagia and opioid-like behavioral depression induced by HT, the albumin fraction (HT) present endogenously in the woodchuck during winter torpor, exerts a direct action on the kidney of the primate, possibly on the mechanisms underlying glomerular filtration and the tubular reabsorption process.

Animals↗

Sympathoadrenal activity and hypoglycemia in the hibernating garden dormouse.

The hibernating garden dormouse is spontaneously hypophagic during the prehibernating period at which time we found a low peripheral sympathetic activity (S.A.). The aim of this work was to investigate the link between dietary intake and S.A. The S.A. was evaluated by measurement of catecholamines in both plasma and adrenal glands by HPLC. Food intake, body weight, energy expenditure and plasma glucose were measured during the reentry phase of the hibernating period. The following results were obtained: the energy intake in pretorpid animals (55 to 83 kJ/24 h/100 g body weight) was less than energy expenditure which was between 145 and 97 kJ/24 h/100 g. The energy deficit induces marked hypoglycemia immediately before the onset of hypothermia (117 mg/dl vs. 76 mg/dl) and leads to a drastic drop in the peripheral sympathetic system. This, in turn, reduced energy production, causing the hypothermia.

Adrenal Glands↗

Characteristics of diaphragm muscle fibre types in hibernating squirrels.

Muscle samples from the diaphragms of 7 full-awake (FA), 10 winter-awake (WA), and 8 hibernating (H) squirrels (Spermophilus lateralis) were quick frozen, sectioned and processed for NADH-TR reaction end-product and myofibrillar-ATPase. Both WA and H squirrels showed small increases in diaphragm weight, reductions in body weight, and hence, significant increases in the diaphragm weight to body weight ratio compared to FA squirrels. They also showed increases in muscle fibre type cross-sectional areas and in the oxidative capacity of type 2b fibres as well as a reduction in capillary density. Furthermore, there also was an increase in the proportion of type 2b fibres in the diaphragm of the H squirrels. Thus, despite the dramatically reduced ventilation associated with hibernation, H squirrels exhibited (1) hypertrophy of the diaphragm which may represent an adaptive response that enables them to work against a stiffer chest wall, and (2) an increased oxidative capacity which enables them to fuel this with fat.

Animals↗

Hibernation hypothermia and metabolism in hedgehogs--changes in free amino acids and related compounds.

1. Hypothermia in midwinter revealed a marked increase in GABA and glutamine due to active decarboxylation and amidation of glutamic acid. This influenced the glutamate-aspartate pathway and resulted in a significant drop in levels of both acids. 2. Elevated levels of GABA and taurine during hibernation pointed to their role as inhibitory neurotransmitters. 3. Amidation of glutamate induced a noticeable drop in ammonia concurring with increased urea and low uric acid levels. 4. Hypothermia in summer revealed a significant role of temperature as a determining factor in the hibernation cycle. Arousal was a repeated, though reversed, phenomenon in this cycle.

Amino Acids↗

In vitro B cell response to glucose in the hibernating hedgehog: comparison with the homeothermic hedgehog and the rat.

The effect of glucose and temperature on insulin secretion was studied using pieces of pancreas from hibernating hedgehogs, homeothermic hedgehogs and rats. The rewarming of the perfusion medium progressively stimulated insulin release from the pancreases from lethargic hedgehogs above 13 degrees C even in the absence of glucose. At low temperature (20 degrees C), insulin probably resulted from labile compartments as suggested by the great first phase of glucose-induced insulin secretion from pancreases from lethargic hedgehogs. The insulin release from pancreases from homeothermic animals (hedgehogs and rats) was temperature dependent only above 23-25 degrees C and only with stimulating glucose concentrations (100 or 300 mg/100 ml). These main differences between B cell physiology of lethargic or homeothermic animals suggest that hibernation induces modifications in the secretory processes which facilitate insulin secretion during the in vivo spontaneous arousal from lethargy.

Animals↗

Arrest of the circadian pacemaker driving the pineal melatonin rhythm in hibernating golden hamsters, Mesocricetus auratus.

Pineal melatonin rhythm in golden hamsters was abolished during hibernation. After arousal in darkness, pineal melatonin increased rapidly regardless of whether the arousal was induced during the day or at night. Rapid increase of pineal melatonin after arousal was markedly diminished in animals exposed to light. In hamsters aroused at midnight, the melatonin rhythm in constant darkness ran with the reversed phase relative to hamsters aroused at noon. Since after arousal the melatonin rhythm obviously starts anew from the same phase, we conclude that the circadian pacemaker driving the rhythm might be arrested during hibernation at the day-time phase.

Animals↗

Hibernation in pygmy possums (Marsupialia: Burramyidae).

The minimum body temperatures and the longest torpor bouts recorded for pygmy possums (family Burramyidae) are compared with hibernating representatives from the insectivores, rodents and Australian bats. The lowest body temperatures of pygmy possums are around 6 degrees C which is within the range of the other mammalian groups. The torpor bouts of the pygmy possums last up to two weeks and the durations are similar to those of eutherian hibernators.

Animals↗