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。     

Seasonal and daily rhythms of body temperature in the European hamster (Cricetus cricetus) under semi-natural conditions

Body temperature of five European hamsters exposed to semi-natural environmental conditions at 47° N in Southern Germany was recorded over a 1.5-year period using intraperitoneal temperature-sensitive radio transmitters. The animals showed pronounced seasonal changes in body weight and reproductive status. Euthermic body temperature changed significantly throughout the year reaching its maximum of 37.9±0.2°C in April and its minimum of 36.1±0.4°C in December. Between November and March the hamsters showed regular bouts of hibernation and a few bouts of shallow torpor. During hibernation body temperature correlated with ambient temperature. Monthly means of body temperature during hibernation were highest in November (7.9±0.8°C) and March (8.2±0.5°C) and lowest in January (4.4±0.7°C). Using periodogram analysis methods, a clear diurnal rhythm of euthermic body temperature could be detected between March and August, whereas no such rhythm could be found during fall and winter. During hibernation bouts, no circadian rhythmicity was evident for body temperature apart from body temperature following ambient temperature with a time lag of 3–5 h. On average, hibernation bouts lasted 104.2±23.8 h with body temperature falling to 6.0±1.7°C. When entering hibernation the animals cooled at a rate of -0.8±0.2°C·h^-1; when arousing from hibernation they warmed at a rate of 9.9±2.4°C·h^-1. Warming rates were significantly lower in November and December than in January and February, and correlated with ambient temperature (r=-0.46, P<0.01) and hibernating body temperature (r=-0.47, P<0.01). Entry into hibrnation occured mostly in the middle of the night (mean time of day 0148 hours ±3.4 h), while spontaneous arousals were widely scattered across day and night. For all animals regression analysis revealed free-running circadian rhythms for the timing of arousal. These results suggest that entry into hibernation is either induced by environmental effects or by a circadian clock with a period of 24 h, whereas arousal from hibernation is controlled by an endogenous rhythm with a period different from 24 h.Abbreviations bw body weight - CET central European time - T _a ambient temperature - T _b body temperature - TTL transistor-transistor logic     

脑室注射6—羟多巴胺对黄鼠科眠入眠的影响

The effect of forced depletion of brain norepinephrine (NE) on the onset of hibernation was observed in the ground squirrel (Citellus dauricus) by intraventricular injection of 6-hydroxydopamine (6-OHDA). The results showed: (1) Intraventricular injection of 100-200 micrograms 6-OHDA, which depleted 50-60% NE, markedly facilitated the onset of hibernation, i.e. the average induction period for hibernation in the treated animals was significantly shorter than that of the natural hibernating animals. (2) The average total torpor time in the treated animals was longer than that of natural hibernating animals. (3) All hibernating animals treated with 6-OHDA were able to wake up from deep hibernation spontaneously and undergo normal hibernation bouts. The results indicate the decrease of NE system activity in brain is one important factor in triggering the onset of hibernation.     

华北地区扬子鳄室内控温冬眠的初步研究

扬子鳄(Alligator sinensis),属于我国Ⅰ级重点保护野生动物,华北地区养殖条件下未见降温冬眠报道.本次于2020年11月至2021年5月对北京动物园饲养的5尾扬子鳄开展人工降温冬眠实验,观察冬眠各期个体的环境选择和行为变化,建立冬眠期间的行为谱,分析冬眠前后行为指标和形态指标(全长、体重和围度)变化,监...     

Age affects hibernation in Syrian hamsters (Mesocricetus auratus)

In this study, we aimed to show how age affects hibernation in the Syrian hamster. Experimentally, we used 30 male animals differing in age. The old animals were 20 months of age and the adults were 8 months of age at the end of the test. The young animals were 3 weeks old at the start of testing and 5 months old at the end of the testing period. The torpor observation started October 15, 1996, and ended March 11, 1997, in the laboratory colony maintained under natural photoperiod and outdoor air. Observations were performed around noon daily. Three measures (i.e., prehibernation period [hibernation latency], proportion of hibernation spent in torpor, and proportion of animals in torpor), all of which reflect the strength of occurrence of hibernation, indicated that the older hamsters (1) started hibernation earlier, (2) spent more time in torpor, and (3) had a higher chance of being in torpor than the younger ones during the hibernation season. (Chronobiology International, 17(5), 623-630, 2000)     

AGE AFFECTS HIBERNATION IN SYRIAN HAMSTERS (MESOCRICETUS AURATUS)

In this study, we aimed to show how age affects hibernation in the Syrian hamster. Experimentally, we used 30 male animals differing in age. The old animals were 20 months of age and the adults were 8 months of age at the end of the test. The young animals were 3 weeks old at the start of testing and 5 months old at the end of the testing period. The torpor observation started October 15, 1996, and ended March 11, 1997, in the laboratory colony maintained under natural photoperiod and outdoor air. Observations were performed around noon daily. Three measures (i.e., prehibernation period [hibernation latency], proportion of hibernation spent in torpor, and proportion of animals in torpor), all of which reflect the strength of occurrence of hibernation, indicated that the older hamsters (1) started hibernation earlier, (2) spent more time in torpor, and (3) had a higher chance of being in torpor than the younger ones during the hibernation season. (Chronobiology International, 17(5), 623–630, 2000)     

Hormonal changes and energy substrate availability during the hibernation cycle of Syrian hamsters

Animals have to adapt to seasonal variations in food resources and temperature. Hibernation is one of the most efficient means used by animals to cope with harsh winter conditions, wherein survival is achieved through a significant decrease in energy expenditure. The hibernation period is constituted by a succession of torpor bouts (hypometabolism and decrease in body temperature) and periodic arousals (eumetabolism and euthermia). Some species feed during these periodic arousals, and thus show different metabolic adaptations to fat-storing species that fast throughout the hibernation period. Our study aims to define these metabolic adaptations, including hormone (insulin, glucagon, leptin, adiponectin, GLP-1, GiP) and metabolite (glucose, free fatty acids, triglycerides, urea) profiles together with body composition adjustments. Syrian hamsters were exposed to varied photoperiod and temperature conditions mimicking different phases of the hibernation cycle: a long photoperiod at 20 °C (LP20 group), a short photoperiod at 20 °C (SP20 group), and a short photoperiod at 8 °C (SP8). SP8 animals were sampled either at the beginning of a torpor bout (Torpor group) or at the beginning of a periodic arousal (Arousal group). We show that fat store mobilization in hamsters during torpor bouts is associated with decreased circulating levels of glucagon, insulin, leptin, and an increase in adiponectin. Refeeding during periodic arousals results in a decreased free fatty acid plasma concentration and an increase in glycemia and plasma incretin concentrations. Reduced incretin and increased adiponectin levels are therefore in accordance with the changes in nutrient availability and feeding behavior observed during the hibernation cycle of Syrian hamsters.     

The effects of cold-stress,hibernation, and prolonged inactivity on bone dynamics in the golden hamster,Mesocricetus auratus

The effects of cold-stress and hibernation on bone dynamics in the femurs of hamsters were investigated using histometric analyses. Control animals were maintained at 27° C for 90 days; experimental animals were kept at 5° C and hibernated for 7, 15, 21, 50, or 90 days. Histometric analyses of cross sections indicated that bone diameter and cortical thickness at the femoral midshaft increased after 83 days of extreme cold and 7 days of hibernation but decreased significantly after 69 days of cold stress and 21 days of hibernation. Osteoporosis was evident although the number of osteons per unit area of bone increased during hibernation. An initial decrease in the number of non-Haversian longitudinal vessels per unit area of bone was seen in experimental animals which was apparently related to a corresponding reduction in cortical thickness. Lacunar area increased in these animals, suggesting that osteocytic osteolysis may be a significant mechanism for calcium regulation during hibernation.     

The impact of cold acclimation and hibernation on antioxidant defenses in the ground squirrel (Spermophilus citellus): An update

Any alteration in oxidative metabolism is coupled with a corresponding response by an antioxidant defense (AD) in appropriate subcellular compartments. Seasonal hibernators pass through circannual metabolic adaptations that allow them to either maintain euthermy (cold acclimation) or enter winter torpor with body temperature falling to low values. The present study aimed to investigate the corresponding pattern of AD enzyme protein expressions associated with these strategies in the main tissues involved in whole animal energy homeostasis: brown and white adipose tissues (BAT and WAT, respectively), liver, and skeletal muscle. European ground squirrels (Spermophilus citellus) were exposed to low temperature (4 ± 1 °C) and then divided into two groups: (1) animals fell into torpor (hibernating group) and (2) animals stayed active and euthermic for 1, 3, 7, 12, or 21 days (cold-exposed group). We examined the effects of cold acclimation and hibernation on the tissue-dependent protein expression of four enzymes which catalyze the two-step detoxification of superoxide to water: superoxide dismutase 1 and 2 (SOD 1 and 2), catalase (CAT), and glutathione peroxidase (GSH-Px). The results showed that hibernation induced an increase of AD enzyme protein expressions in BAT and skeletal muscle. However, AD enzyme contents in liver were largely unaffected during torpor. Under these conditions, different WAT depots responded by elevating the amounts of specific enzymes, as follows: SOD 1 in retroperitoneal WAT, GSH-Px in gonadal WAT, and CAT in subcutaneous WAT. Similar perturbations of AD enzymes contents were seen in all tissues during cold acclimation, often in a time-dependent manner. It can be concluded that BAT and muscle AD capacity undergo the most dramatic changes during both cold acclimation and hibernation, while liver is relatively unaffected by either condition. Additionally, this study provides a basis for further metabolic study that will illuminate the causes of these tissue-specific AD responses, particularly the novel finding of distinct responses by different WAT depots in hibernators.     

Further studies on opioids and hibernation: delta opioid receptor ligand selectively induced hibernation in summer-active ground squirrels

To examine the possible involvement of multiple opioid receptors in animal hibernation, we infused opioids selective for mu, kappa, and delta opioid receptors into summer-active ground squirrels (Citellus tridecemlineatus). The effects of those opioid treatments on the hibernation induced by HIT (Hibernation Induction Trigger) were also examined. Mu opioids morphine (1.50 mg/kg/day) and morphiceptin (0.82 mg/kg/day) and kappa opioid peptide dynorphin A (0.82 mg/kg/day) did not induce hibernation. On the contrary, morphine, morphiceptin and dynorphin A antagonized HIT-induced hibernation in summer-active ground squirrels. Infusion of delta opioid DADLE (D-Ala2-D-Leu5 enkephalin; 1.50 mg/kg/day), however, induced summer hibernation in a manner comparable to that induced by HIT. It is concluded therefore that delta opioid receptor and its ligand may be intimately involved in animal hibernation. In view of the fact that HIT was obtained from winter hibernating animals and might therefore be responsible for natural hibernation, our results also suggest that naturally occurring mu and kappa opioids may play an important role in the arousal state of hibernation.     

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.     

Photoperiodic regulation of body mass, food intake, hibernation, and reproduction in intact and castrated male European hamsters,Cricetus cricetus

A group of sexually active male European hamsters were raised either in short-photoperiod conditions (SP; LD 8:16) or in long-photoperiod conditions (LP; LD 16:8) from their capture at the end of the hibernation period. Another group of hamsters was castrated in April and gonadectomized animals were maintained in SP and cold (7 degrees C) or in a succession of SP and LP plus cold. Another group, castrated in May or in September and raised in LP conditions, was transferred in September to SP conditions and cold. 1. Normal hamsters raised in continuous SP or LP apparently did not show signs of rhythmic behavior, except possibly in gonadal activity. 2. Body weight increased continuously, plasma testosterone levels oscillated between 1.5 and 2.5 ng/ml, and animals raised in SP and in cold did not enter hibernation. 3. Similar results were also found in castrated animals kept in SP conditions and cold. 4. The sequence LP-SP induced a decrease in food intake and body weight and a decrease in plasma testosterone levels and triggered entry into hibernation in both intact and castrated animals. 5. After 6 months continuously in SP and with exposure to cold spontaneous recrudescence in food intake and body weight occurred and hibernation ended in both intact and castrated animals. 6. In normal animals a spontaneous increase in plasma testosterone levels was observed. 7. In both normal and gonadectomized animals the phase of refractoriness could be broken by exposure to LP conditions. 8. The critical photoperiod lies between 15 and 15.5 h. These results demonstrate that the European hamster is a photoperiodic species.(ABSTRACT TRUNCATED AT 250 WORDS)     

Testis development and testosterone secretion in captive European ground squirrels before, during, and after hibernation

In some hibernating species, an extended euthermic period before spring emergence has been reported during which testicular maturation occurred. In this study, we investigated whether male European ground squirrels Spermophilus citellus (Linnaeus, 1766) develop scrotal testes before or after the termination of hibernation. The course of testis development and testosterone concentrations were examined in young-of-year male ground squirrels (n = 4) before, during, and after their first hibernation. In the environmental chamber, all animals showed regular hibernation patterns with decreasing torpor bout lengths prior to the end of hibernation. Four weeks before hibernation ended, testosterone levels were elevated during spontaneous arousals. Testosterone concentrations peaked during the first 2 months post heterothermy and decreased thereafter. In 2 experimental males, testes descended from the inguinal region into the scrotum shortly before the end of heterothermy. The state of testis development and testosterone secretion corresponded to that found in semi-free-living males at spring emergence. Testis size increased in active animals to maximum widths during weeks 3 and 4 post heterothermy. Scrotal skin pigmentation was absent until heterothermy ended and developed during the first month thereafter. The data suggest that male S. citellus do not need a pre-emergence euthermic period for reproductive development.     

Levels of gap junction proteins in coronary arterioles and aorta of hamsters exposed to the cold and during hibernation and arousal.

There are marked changes in vascular dynamics during prolonged periods in the cold, entrance into hibernation, and arousal to euthermy. Cell-to-cell communication through gap junction channels plays a pivotal role in the control of vasomotor function. Multiple gap junction proteins are expressed in blood vessels, including connexins 37 (Cx37), 40 (Cx40), 43 (Cx43), and 45 (Cx45). Using immunolabeling techniques combined with confocal microscopy, we quantitated the levels of these connexins in coronary arterioles and the thoracic aorta of the golden hamster in four physiological conditions: normal control animals at euthermy; cold-exposed animals (before entrance into hibernation); during hibernation; and after 2-hr arousal from hibernation. In all groups, Cx37 was localized between endothelial cells of the aorta and Cx40 was observed between endothelial cells of coronary arterioles and the aorta. Cx43 was confined to smooth muscle cells of the aorta. Labeling for Cx45 was detected in the endothelium of the ascending aorta. The expression of Cx37 was significantly reduced in cold-exposed, hibernating, and aroused animals. Immunolabeling for Cx40 was increased in the coronary arteriolar endothelium of the cold-exposed group compared with normal controls, hibernating, and aroused animals, perhaps to facilitate intercellular communication during the prolonged circulatory changes to vascular dynamics required to maintain core temperature during cold adaptation. Cx40 expression was unchanged in the aorta. Cx43 immunoexpression in the aorta remained constant under all conditions examined. These changes in connexin expression did not occur during the rapid circulatory changes associated with arousal from hibernation.     

The brain 5-HT1A receptor gene expression in hibernation

Hibernation is a unique physiological state characterized by profound reversible sleep-like state, depression in body temperature and metabolism. The serotonin 5-hydroxytryptamine_1A (5-HT_1A) receptor gene sequence in typical seasonal hibernator, ground squirrel ( Spermophilus undulatus ), was specified. It was found that the fragment encoding the fifth transmembrane domain showed 93.6% of homology with the analogous fragment of the mouse and rat genes and displayed 88.5% homology with the human 5-HT_1A receptor gene. Using primers designed on the basis of obtained sequence, the expression of 5-HT_1A receptor gene in the brain regions in active, entering into hibernation, hibernating and coming out of hibernation ground squirrels was investigated. Significant structure-specific changes were revealed in the 5-HT_1A messenger RNA (mRNA) level in entry into hibernation and in arousal. An increase in the 5-HT_1A gene expression was found in the hippocampus during the prehibernation period and in ground squirrels coming out of hibernation, thus confirming the idea of the hippocampus trigger role in the hibernation. Significant decrease in 5-HT_1A receptor mRNA level in the midbrain was found in animals coming out of hibernation. There was no considerable changes in 5-HT_1A receptor mRNA level in different stages of sleep–wake cycle in the frontal cortex. Despite drastically decreased body temperature in hibernating animals (about 37°C in active and 4–5°C in hibernation), 5-HT_1A receptor mRNA level in all examined brain regions remained relatively high, suggesting the essential role of this 5-HT receptor subtype in the regulation of hibernation and associated hypothermia.     

Lactation during hibernation in wild black bears: effects on plasma amino acids and nitrogen metabolites.

This study examined the seasonal and reproductive influences on individual plasma amino acid concentrations and nitrogen metabolites in a black bear population (Ontario, Canada). During hibernation, 11 of 23 plasma amino acids were significantly higher (13%-108%) in lactating than in nonlactating females, without an alteration in plasma total protein or total essential or nonessential amino acid levels. The greatest changes were observed in glutamine, arginine, and glycine levels. Plasma urea, urea/creatinine, and ammonia levels were significantly lower in hibernating compared with active female bears, but lactation had no effect on these parameters. Taken together these results show that lactation during hibernation is an additional metabolic challenge that results in increased mobilization of individual plasma amino acids and no accumulation of nitrogen end products, underlining the remarkable efficiency of amino acid and urea recycling in denning female black bears.     

Hibernation does not affect memory retention in bats

Long-term memory can be critically important for animals in a variety of contexts, and yet the extreme reduction in body temperature in hibernating animals alters neurochemistry and may therefore impair brain function. Behavioural studies on memory impairment associated with hibernation have been almost exclusively conducted on ground squirrels (Rodentia) and provide conflicting results, including clear evidence for memory loss. Here, we for the first time tested memory retention after hibernation for a vertebrate outside rodents—bats (Chiroptera). In the light of the high mobility, ecology and long life of bats, we hypothesized that maintenance of consolidated memory through hibernation is under strong natural selection. We trained bats to find food in one out of three maze arms. After training, the pre-hibernation performance of all individuals was at 100 per cent correct decisions. After this pre-test, one group of bats was kept, with two interruptions, at 7°C for two months, while the other group was kept under conditions that prevented them from going into hibernation. The hibernated bats performed at the same high level as before hibernation and as the non-hibernated controls. Our data suggest that bats benefit from an as yet unknown neuroprotective mechanism to prevent memory loss in the cold brain.     

达乌尔黄鼠实验室饲养、繁殖及其冬眠阵

为探索实验室条件下达乌尔黄鼠饲养与繁殖的方法及冬眠阵的发生规律,参照野生黄鼠冬眠洞穴的主要生态环境参数,建立人工冬眠屋,采用传统锯末技术记录冬眠阵。结果显示: (1) 处于春季繁殖期的黄鼠应以大鼠饲料为主,辅以少量黄瓜等,夏季活跃期交叉饲喂大鼠饲料与兔饲料,辅以多水的瓜果蔬菜,秋季育肥期以大鼠饲料为主,辅以高脂肪高蛋白的花生、豆类等。(2)雌鼠怀孕期为28 d 左右,哺乳期约一个月,雌鼠每窝产仔4 ～ 8 只,平均5.52 只;初生幼鼠两周内忌换垫料,并避免将异味带入鼠房。(3)黄鼠冬眠期从当年11月下旬至次年3 月上旬,平均93.95 d;冬眠阵睡眠时长平均7. 44 d,阵间激醒时长平均1.36 d,睡眠天数占整个冬眠期的89.9% ;整个冬眠期,黄鼠冬眠阵平均7. 55 个。(4)2009 年秋至2011 年春季,自野外共捕回黄鼠185 只, 存活146 只,存活率78. 9% 。在2006、2009 和2011 年的黄鼠繁殖期,共配对25 对,产仔138 只,成活92 只,成活率为66.7% 。结果表明,野生达乌尔黄鼠可在人工饲养条件下实现繁殖,并可在人工冬眠屋成功冬眠。     

Hibernation and disuse osteoporosis.

In an attempt to determine 1) if disuse osteoporosis occurs naturally during hibernation and 2) if it can be induced by limb immobilization in active and hibernating ground squirrels, forty-six thirteenlined ground squirrels were divided into five experimental groups. Group one was the active control group (AC) (ten animals). Group two was the active immobilized (AI) group and was subjected to a unilateral sciatic nerve section (ten animals). Group three was the hibernating control group and was allowed to hibernate undisturbed for 30 days (HC) (ten animals). The fourth group was subjected to a unilateral sciatic nerve section and allowed to hibernate for 30 days; this was the hibernating immobilized group (HI). The fifth group of six animals received the identical treatment as the (HI) group but was allowed to hibernate for 150 days.The following measurements were taken: Gastrocnemius-soleus mass, body mass, tibia-fibula weight, total tibia-fibula calcium and phosphorus. In addition, the femur was observed histologically for the enlarged lacunae typical of osteoporotic bone. Statistical analysis was accomplished with the Wilcoxan sign rank test.The data were interpreted to mean: 1) Disuse osteoporosis can be induced in active ground squirrels, 2) disue osteoporosis is not evident nor can it be induced by limb immobilization during 30 days of hibernation and 3) limb immobilization did not result in significant disuse osteoporosis following 150 days of hibernation. Only a few enlarged lacunae were evident after 150 days of hibernation.