Age-class differences in the pattern of hibernation in yellow-bellied marmots,Marmota flaviventris

Summary Age-related differences in the patterns of body temperature regulation during hibernation were found in yellow-bellied marmots. The timing of all entrances into and arousals from torpor was determined from continuous records of thermocouples mounted in each animal's nest box. Older marmots spent more time at high body temperatures following periodic arousals from torpor than did juveniles undergoing their first season of hibernation. In addition, older marmots spontaneously terminated their hibernation seasons in the spring, whereas most juveniles continued to hibernate until either they were emaciated from starvation or they were fed. These two patterns of hibernation reflect age- and size-related differences in the degree to which the animals are constrained energetically and the probability that they can successfully reproduce in spring. The patterns also are consistent with age-related differences in the timing of dormancy in nature.     

Oxygen consumption and body temperature in yellow-bellied marmot populations from montane-mesic and lowland-xeric environments

Summary Yellow-bellied marmots characteristically live in montane-mesic environments, but in several areas in western North America, this species extended its range into lowland-xeric habitats. Body mass was significantly smaller in the lowland-xeric population from eastern Washington at 393 m than in the montane-mesic population from western Colorado at 2900 m. Oxygen consumption of marmots from montane-mesic and lowland-xeric environments was signiflcantly affected by ambient temperature (T_A) water regimen, population, and a population x water regimen x temperature interaction. Lowland-xeric animals had a higher metabolic rate at low T_As, but a lower metabolic rate at higher T_As than the montane-mesic aminals. Oxygen consumption was lower on a restricted-water regimen than on ad libitum water in both populations. Coefficients relating oxygen consumption to body mass were affected by T_A, water regimen, and population. These intraspecific coefficients are larger than the interspecific coefficients for all mammals. Body temperature (T_B) was affected significantly by T_A, water regimen, and population. T_A body mass, and a population x water regimen interaction significantly affected conductance. Conductance generally was higher in the lowland-xeric than in the montane-mesic marmots. Both populations increased conductance at high T_A, but the lowland-xeric population dissipated a much higher proportion of the heat by evaporative water loss (EWL) than did the montane-mesic population. Metabolic water production exceeded or equaled EWL at 5–20°C. Smaller body size, reduced metabolism at high T_A, and increased EWL at high T_A characterized the lowland-xeric population.Metabolic rates of yellow-bellied marmots were higher than predicted from body size during the reproductive season but decreased to 67% of that predicted from the Kleiber curve by late summer. Marmots minimize thermoregulatory costs by concentrating activity at times when the microclimate is favorable, by tolerating hyperthermia at high T_A in the field, and by having a conductance lower than that predicted from body size.Abbreviations DHC dry-heat conductance - EHL evaporative heat loss - EWL evaporative water loss - HP heat produced - T _A ambient temperature - T _n body temperature - M body mass     

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.     

Seasonal changes in serum metabolites in free-ranging alpine marmots (<Emphasis Type="Italic">Marmota marmota</Emphasis>)

Circannual changes in serum parameters and body mass were studied in free-ranging Marmota marmota that had been shot in Switzerland (Grisons) for management reasons in May, July, and September of 1995, 1996, and 1997; and in April 1996. Markers of lipid (triglycerides, cholesterol), protein (total protein, urea-nitrogen) and mineral (calcium, inorganic phosphate) metabolism were evaluated in 111 haemolysis-free serum samples; the effects of sex, age, reproductive status, season and year were tested. Mean body mass was higher in adult males than in adult females in July and September, and serum cholesterol concentrations were lower in adult males in May. Pregnant females had lower concentrations of total protein than non-pregnant females, and triglyceride concentrations were negatively correlated with the number of uterine ampullae. Inorganic phosphate decreased and total protein increased with age. In adults, triglycerides, cholesterol and urea-nitrogen increased mainly from May to July and decreased between September and May. Total protein, calcium and phosphate did not change throughout the year. This suggests that, during the active season, ingested lipids and amino acids were utilised for metabolic needs or lipid storage, while in winter, lipids were catabolised and protein was spared. Quantitative differences between years, observed for triglycerides and total protein during the active season, were probably due to different climatic conditions in each year.     

Effect of predation risk on the presence and persistence of yellow-bellied marmot (Marmota flaviventris) colonies

Habitat selection may have population level consequences and ultimately may influence a population's local persistence or extinction. We capitalized on a long-term study (1962–2004) of yellow-bellied marmots Marmota flaviventris in and around the Rocky Mountain Biological Laboratory, Gothic, Colorado, USA, and compared habitat characteristics associated with food availability and predation risk to explain variation in persistence of marmots at 27 sites, and their absence at 22 additional, randomly selected sites. We classified sites as persistent, intermittent or null based on whether there was a history of extinction; intermittent sites periodically went extinct and null sites never had marmots. Logistic regression analyses revealed that environmental variables associated with visibility and safety, rather than food, correctly classified sites as persistent or non-persistent as well as persistent or intermittent. Discriminant function analysis that included the null sites revealed that the same visibility-related characteristics predicted where marmots were found. These results highlight the importance of variation in safety among sites in predicting long-term population persistence, as well as a species' distribution.     

Genetic diversity-fitness correlation revealed by microsatellite analyses in European alpine marmots (Marmota marmota)

The relationship between individual genetic diversity and fitness-related traits are poorly understood in the wild. The availability of highly polymorphic molecular markers, such as microsatellites, has made research on this subject more feasible. We used three microsatellite-based measures of genetic diversity, individual heterozygosity H, mean d ² and mean d ² _outbreeding to test for a relationship between individual genetic diversity and important fitness trait, juvenile survival, in a population of alpine marmots (Marmota marmota), after controlling for the effects of ecological, social and physiological parameters that potentially influence juvenile survival in marmots. Analyses were conducted on 158 juveniles, and revealed a positive association between juvenile survival and genetic diversity measured by mean H. No association was found with mean d ² and with mean d ² _outbreeding. This suggests a fitness disadvantage to less heterozygous juveniles. The genetic diversity-fitness correlation (GDFC) was somewhat stronger during years with poor environmental conditions (i.e. wet summers). The stressful environmental conditions of this high mountain population might enhance inbreeding depression and make this association between genetic diversity and fitness detectable. Moreover the mating system, allowing extra pair copulation by occasional immigrants, as well as close inbreeding, favours a wide range of individual genetic diversity (mean H ranges from 0.125 to 1), which also may have facilitated the detection of the GDFC. The results further suggest that the observed GDFC is likely to be explained by the “local effect” hypothesis rather than by the “general effect” hypothesis.     

Seasonal changes in ultrastructure of brown adipose tissue in the common shrew (Sorex araneus L.)

Summary Seasonal changes have been detected in the ultrastructure of brown adipose tissue (BAT) of the common shrew, Sorex araneus (a true non-hibernator) living under natural conditions and collected at the time when the most representative growth phase of the animal for the given season could be expected.In summer and autumn, BAT is characterized by the presence of large, regular, spherical lipid droplets and mitochondria closely adhering to one another.During winter, mitochondria possess densely packed cristae and are dispersed in the cytoplasm, sometimes invaginating into lipid droplets; the latter are diminished and often irregular in contour. The BAT in winter specimens is distinguished also by a large amount of blood capillaries penetrating the tissue.In spring, mitochondria of BAT are found more frequently adhering to each other, and are characterized by loosely arranged cristae. In addition to the spherical lipid droplets, agglomerations of lipid material may be found in the cytoplasm.The observed seasonal fluctuations in the ultrastructure of BAT in the shrew correspond to the metabolic rhythm of this animal. The latter point is discussed.The author wishes to thank Prof. Zdzisaw Pucek, Head of Mammal Research Institute in Biaowiea, for valuable discussions on the biology of shrews and for providing all the necessary facilities for carrying out this study.The skillful technical assistance of Mrs. Z. Karunos and Mrs. K. Mroziska is gratefully acknowledged     

Seasonal changes of thermogenic capacity in Melano-bellied oriental voles (Eothenomys melanogaster)☆

Seasonal changes of thermogenic capacity in Melano-bellied oriental voles (Eothenomysmelanogaster) were studied by measurements of body mass, basal metabolic rate (BMR), non-shivering thermogenesis (NST), thermogenic properties of brown adipose tissue (BAT), relative weight of liver mass, mitochondrial protein (MP) content, and activities of mitochondrial cytochrome C oxidase (COX) in autumn (October), winter (December), spring (April) and summer (July), respectively.
Results showed that: (1) Body mass of E.melanogaster was the highest in autumn, and the lowest in summer. The seasonal change in body mass was subject not only by environment temperature but also to other factors such as reproduction, feeding habit and climate in the habitat. Reproductive periods of E.melanogaster were in early spring (February–March) and late autumn (September–October), and therefore, the body mass was higher in spring and autumn. In addition, E.melanogaster feeds on leaves and stems of plants in spring and summer, whereas mainly on fruits in autumn and winter as they contain more energy. The difference in the feeding habit could result in higher body mass in the late half of the year, but lower body mass in winter. The lower body mass in winter provides E.melanogaster with an advantage in motility. (2) The BMR and NST of E.melanogaster was higher in winter than in other seasons, which suggests that the regulating thermogenic capacity is an important adaptive strategy for E.melanogaster. However, there was no difference in relative oxygen consumption (NST–BMR/BMR), suggesting that the regulative capacity of BMR is approximately equal to NST in E.melanogaster. This is different from some small mammal species in north China, which have higher regulative capacity in NST than E.melanogaster. Melano-bellied oriental voles live in mountainous area with subtropical zone monsoon climate and abundant food resource, and therefore have lower regulative capacity in NST, which can reduce an absolute request for energy. The low regulative capacity in NST can also help E.melanogaster to maintain energy balance and stable body temperature. (3) The relative mass of BAT in winter was significantly higher than that in summer. Different seasons significantly affected the MP content of BAT. The MP content of BAT was in the following order: winter > fall > spring > summer. The trend as following was observed for the mitochondrial COX activity of BAT: winter > spring > fall > summer. In experimental conditions, the thermogenic rate of many small mammal species increased under cold acclimation. Short photoperiod alone could also induce E.melanogaster to increase NST and mitochondrial COX activity of BAT. Therefore, ambient temperature and photoperiod are important environmental cues to induce seasonal changes in thermogenic capacity in E.melanogaster. (4) The relative mass of liver in summer was higher than that in other seasons, but no seasonal changes were found among other seasons. The MP contents of liver showed highest level in winter and lowest in spring and summer. Significant seasonal changes were also observed in the mitochondrial COX activity of liver. It increased significantly in winter, but not in other seasons. This indicates that liver participate in adjustment of thermogenesis. These data indicate that both increasing whole-animal thermogenesis rate and thermogenic capacity at the cells of BAT and liver play an important role in the adaption of E.melanogaster to seasonal changes, and they are also closely correlated with the habitat condition and their life habit.     

Seasonal changes of thermogenic capacity in Melano-bellied oriental voles (Eothenomys melanogaster)

Seasonal changes of thermogenic capacity in Melano-bellied oriental voles (Eothenomysmelanogaster) were studied by measurements of body mass, basal metabolic rate (BMR), non-shivering thermogenesis (NST), thermogenic properties of brown adipose tissue (BAT), relative weight of liver mass, mitochondrial protein (MP) content, and activities of mitochondrial cytochrome C oxidase (COX) in autumn (October), winter (December), spring (April) and summer (July), respectively.
Results showed that: (1) Body mass of E.melanogaster was the highest in autumn, and the lowest in summer. The seasonal change in body mass was subject not only by environment temperature but also to other factors such as reproduction, feeding habit and climate in the habitat. Reproductive periods of E.melanogaster were in early spring (February–March) and late autumn (September–October), and therefore, the body mass was higher in spring and autumn. In addition, E.melanogaster feeds on leaves and stems of plants in spring and summer, whereas mainly on fruits in autumn and winter as they contain more energy. The difference in the feeding habit could result in higher body mass in the late half of the year, but lower body mass in winter. The lower body mass in winter provides E.melanogaster with an advantage in motility. (2) The BMR and NST of E.melanogaster was higher in winter than in other seasons, which suggests that the regulating thermogenic capacity is an important adaptive strategy for E.melanogaster. However, there was no difference in relative oxygen consumption (NST–BMR/BMR), suggesting that the regulative capacity of BMR is approximately equal to NST in E.melanogaster. This is different from some small mammal species in north China, which have higher regulative capacity in NST than E.melanogaster. Melano-bellied oriental voles live in mountainous area with subtropical zone monsoon climate and abundant food resource, and therefore have lower regulative capacity in NST, which can reduce an absolute request for energy. The low regulative capacity in NST can also help E.melanogaster to maintain energy balance and stable body temperature. (3) The relative mass of BAT in winter was significantly higher than that in summer. Different seasons significantly affected the MP content of BAT. The MP content of BAT was in the following order: winter > fall > spring > summer. The trend as following was observed for the mitochondrial COX activity of BAT: winter > spring > fall > summer. In experimental conditions, the thermogenic rate of many small mammal species increased under cold acclimation. Short photoperiod alone could also induce E.melanogaster to increase NST and mitochondrial COX activity of BAT. Therefore, ambient temperature and photoperiod are important environmental cues to induce seasonal changes in thermogenic capacity in E.melanogaster. (4) The relative mass of liver in summer was higher than that in other seasons, but no seasonal changes were found among other seasons. The MP contents of liver showed highest level in winter and lowest in spring and summer. Significant seasonal changes were also observed in the mitochondrial COX activity of liver. It increased significantly in winter, but not in other seasons. This indicates that liver participate in adjustment of thermogenesis. These data indicate that both increasing whole-animal thermogenesis rate and thermogenic capacity at the cells of BAT and liver play an important role in the adaption of E.melanogaster to seasonal changes, and they are also closely correlated with the habitat condition and their life habit.     

Seasonal glucose uptake in marmots (Marmota flaviventris): the role of pancreatic hormones.

1. Glucose uptake was measured throughout the year in marmots (Marmota flaviventris) by the hyperglycemic clamp technique. During each 2 hr experiment, the plasma glucose level was maintained at 215 mg/dl while blood samples were collected and analysed for glucose, insulin, glucagon, cortisol and catecholamines. 2. Glucose uptake was calculated from the glucose infusion rate, changes in the glucose pool (using a correction factor), and urinary glucose excretion. 3. In autumn, animals peaked in body weight (greater than 4.0 kg) and ceased to feed. Basal plasma insulin levels in autumn were significantly elevated over all other seasons (P less than 0.01) and glucose uptake in autumn was 9.7 +/- 2.4 mg/min which was significantly lower (P less than 0.05) than summer (21.7 +/- 2.4 mg/min) during the steady state phase of the glucose clamp (90-120 min). 4. Plasma glucagon levels declined during the clamp in all seasons but there was no significant difference between seasons. Plasma cortisol and catecholamine (norepinephrine and epinephrine) levels remained unchanged under basal and experimental conditions throughout the seasons. 5. During glucose infusion, beta-hydroxybutyrate levels were suppressed suggesting that lipolysis was reduced during the experiment. 6. These results suggest that the marmot exhibits seasonal changes in glucose uptake; the lowest rate of glucose uptake occurring in the autumn after the animals peak in body weight and cease to feed.     

Decreased rate of fatty acid synthesis in brown adipose tissue of hypothalamic obese rats

Intranuclear coinjection of the late SV40 KpnI/BclI DNA fragment and the early promotor/enhancer HpaII/BglI DNA segment into permissive monkey and non-permissive mouse cells allows late SV40 gene expression without T-antigen synthesis and DNA replication. These conditions were chosen to analyse the effect of DNA methylation on V-antigen synthesis detached from the process of DNA replication. We found that in vitro methylation of a single cytosine nucleotide proximal to the major late mRNA cap site by the HpaII methylase does not block capsid protein synthesis. This result is in contrast to reported data obtained in Xenopus laevis oocyte injection experiments [(1982) Proc. Natl. Acad. Sci. USA 79, 5142-5146].     

Microsatellite variation in autochthonous and introduced populations of the Alpine marmot (Marmota marmota) along a European west–east transect

Microsatellite variation was studied in 11 populations of the Alpine marmot along a west–east transect through the present distribution range. The samples represent five autochthonous and six introduced populations. Eleven loci were analysed in nine populations and six loci in the two populations from France. In the populations from the Western Alps, there is no indication for reduced variability as has been assumed in previous studies. However, a decrease of variation in the autochthonous populations was observed from the west to the east. The introduced populations showed a heterogeneous pattern reflecting the geographic origin of the released individuals. The population from the Spanish Pyrenees harbours a high level of variation and is genetically closest to the French populations. In Austria, three of the introduced populations have low variation and are closely related to the autochthonous populations from the western part of Austria. In contrast, two introduced populations from the central part of Austria are highly variable and resemble the populations from France. At least for one of these populations an early introduction of founder individuals from the Western Alps has been documented.     

Metabolism,thermogenesis and daily rhythm of body temperature in the wood lemming,Myopus schisticolor

Wood lemmings (Myopus schisticolor) were captured during their autumnal migration in September and October. The animals were maintained at 12°C and under 12L:12D photoperiod. Basal metabolic rate and thermogenic capacity of the wood lemming were studied. Basal metabolic rate was 3.54 ml O₂·g^-1·h^-1, which is 215–238% of the expected value. The high basal metabolic rate seems to be typical of rodents living in high latitudes. The body temperature of the wood lemming was high (38.0–38.8°C), and did not fluctuate much during the 24-h recording. The high basal metabolic rate and the high body temperature are discussed with regard to behavioural adaptation to a low-quality winter diet. Thermogenic capacity, thermal insulation and non-shivering thermogenesis of the wood lemming displayed higher values than expected: 53.0 mW·g^-1, 0.53 mW·g^-1·C^-1 and 53.2 mW·g^-1, respectively. Brown adipose tissue showed typical thermogenic properties, although its respiratory property was fairly low, but mitochondrial protein content was high compared to other small mammals. The 24-h recording of body temperature and motor activity did not reveal whether the wood lemming is a nocturnal animal. Possibly, the expression of a circadian rhythm was masked by peculiar feeding behaviour. It is concluded that the wood lemming is well adapted to living in cold-temperature climates.Abbreviations BAT brown adipose tissue; bm, body mass - BMR basal metabolic rate - C conductance - Cox cytochrome-c-oxidase - HP heat production - HP_max maximum heat production - M metabolism - NA noradrenaline - NST non-shivering thermogenesis - NST_max maximum non-shivering thermogenesis - RMR resting metabolic rate - RQ respiratory quotient - T _a anibient temperature - T _b body temperature - T _lc lower critical temperature - UCP uncoupling protein - vO₂ oxygen consumption - vO_{2 max} maximum oxygen consumption     

Effects of high fat diets on hibernation and adipose tissue in Turkish hamsters

Summary The effects of dietary fat saturation and fat content on hibernation and several properties of white and brown adipose tissue (WAT and BAT, respectively) were investigated in Turkish hamsters (Mesocricetus brandti). Male hamsters were housed in a long photoperiod (LD 16:8) at 23°C and fed one of three diets: (1) chow (6.5% fat per weight), (2) chow+13.5% vegetable oil (OIL, 20% fat per weight [largely unsaturated fat]) and (3) chow+13.5% vegetable shortening [SHORTENING, 20% fat per weight (largely saturated fat)]. Five weeks later body weights had stabilized and the animals were transferred to a short photoperiod (LD 8:16) at 3°C. At the peak of the hibernation season (17 weeks) the animals were sacrificed within 24 h of arousal. Chow-fed hamsters had the greatest percentage of animals hibernating and days found torpid compared with the two fat-fed groups, with no differences found between the latter two groups for these measures. There were no differences between hibernating (HIB) and nonhibernating (NON-HIB) hamsters across or within the diet groups for any of the BAT measures [uncoupling protein content, mitochondrial mass, lipoprotein lipase (LPL) activity, and in vivo lipogenesis], nor were there significant effects of the diet on these measures. CHOW-and OIL-fed HIB hamsters showed decreases in body weight. All HIB groups had decreases in each carcass component, several fat pad weights, testes weight, and food intake. No consistent differences in WAT LPL activity or in vivo lipogenesis were found between HIB and NON-HIB hamsters. Feeding saturated high fat diets inhibits hibernation in some species; however, in the present experiment, feeding of both saturated and unsaturated fat-laden diets inhibited hibernation to a similar degree.Abbreviations BAT brown adipose tissue - COA cytochrome-c oxidase - DS dorsal subcutaneus - DSWAT dorsal subcutaneous white adipose tissue - E epididymal - EWAT epididymal white adipose tissue - FFDM fat-free dry mass - HIB hibernating - I interscapular - IBAT intercapsular brown adipose tissue - IS inguinal subeutaneus - ISWAT inguinal subcutaneous white adipose tissue - LPL lipoprotein lipase - NON-HIB non-hibernating - R retroperitoneal - RWAT retroperitoneal white adipose tissue - SDS sodium dodecyl sul - UCP uncoupling protein - WAT white adipose tissue     

Maintenance of biological rhythms during hibernation in Eastern woodchucks (<Emphasis Type="Italic">Marmota monax</Emphasis>)

We undertook a study to determine presence of circadian rhythms during woodchuck hibernation using continuously monitored body temperatures. Males had shorter torpor and longer euthermic periods than females. Circular statistics revealed a significant mean vector for males entering into torpor (10:21 h), but not for females. No significant mean vector was found for male or female arousal from torpor. A contingency test was applied to the torpor bout durations. All 7 males tested had significant τ’s between 24 and 26 h, while 6 of the 13 females tested had significant τ’s with a range of 22–27 h. These results implicate a free-running circadian clock during torpor bouts. Overall, the data support the existence of biological rhythms during hibernation in woodchucks, especially for males during arousals. Since entries into torpor appear to be synchronized for males, arousal periods may be used to resynchronize their circadian system. The persistence of biological rhythms during hibernation may help to insure successful mating in the spring after emergence.     

Long photophase is not a sufficient stimulus to reduce thermogenic capacity in winter-acclimatized short-tailed field voles (Microtus agrestis) during long-term cold acclimation

The thermogenic capacity of brown adipose tissue in winter- and summer-acclimatized short-tailed field voles (Microtus agrestis) was investigated by examining changes in mass of brown adipose tissue, the ratio of white adipose tissue to brown adipose tissue, the concentration of the uncoupling protein (thermogenin) in whole depots (μg) and in mitochondrial mass (μg·mg^-1) and the activity of cytochrome c oxidase in the depots (mmol·min^-1). The concentration of thermogenin in winter-acclimatized voles (n=8), per brown adipose tissue depot and per mitochondrial mass, was significantly higher than in summer-acclimatized voles (n=6). There was no significant difference in the level of cytochrome c oxidase activity between these two groups. Four groups of winter-acclimatized voles (n=6 in each group) were exposed to 5°C for 10, 20, 50 and 100 days in a 14L:10D photoperiod. Body mass, brown adipose tissue mass, white adipose tissue mass and basal metabolic rate were significantly positively related to the length of time cold exposed up to 100 days. There was a significant inverse relationship between the ratio of white to brown adipose tissue mass and the duration of cold exposure. There was no significant relationship between thermogenin concentration, either per depot or in mitochondrial mass of brown adipose tissue, with the length of time cold exposed. The level of cytochrome c oxidase activity increased significantly from control levels to a maximum after 10 days in the cold but decreased from 10 days onwards. In winter-acclimatized M. agrestis, a 14L:10D photoperiod is not a sufficient stimulus to reduce thermogenic capacity during cold acclimation. Indeed, some changes in the indirect parameters reflecting thermogenesis, notably the increase in basal metabolic rate and the decrease in the ratio of white to brown adipose tissue mass, indicated that despite the long photophase the thermogenic capacity was slightly further enhanced during the cold acclimation.     

Seasonal changes in hepatocyte ultrastructure correlated with the cyclic synthesis of secretory proteins in the winter flounder (Pleuronectes americanus)

Liver tissue was sampled from flounder (Pleuronectes americanus) throughout the year with the intention of documenting changes in the ultrastructure coincident with the production and secretion of antifreeze proteins. In the winter, hepatocytes are dedicated to the production of these proteins and, in the female, also reproductive proteins. In both sexes, liver cells in the summer contain abundant lipid and glycogen stores. In the female, there is a conspicuous hepatocyte transformation from a fat-filled cell in the summer to one with well-developed rough endoplasmic reticulum in the winter. Large amounts of rough endoplasmic reticulum (11.2 mg/gm) were recovered after subcellular fractionation of female wintertime liver. The increased appearance of secretory organelles and the high number of nucleolar profiles observed in winter animals is consistent with the elevated demand for protein secretion and synthesis in both sexes. The fractional volumes occupied by lipid droplets and mitochondria were different when comparisons were made between sex and season. Females contained a greater volume of lipid than did males, and summer animals contained more lipid than those in winter.     

Brown fat temperature decrease by electrical stimulation of in and around retrorubral field in the golden hamster

Electrical stimulation (0.1 ms, 1 mA, 5–50 Hz) of the ventral midbrain of the golden hamster decreased frequency-dependent temperatures of the interscapular brown adipose tissue (IBAT) and rectum. The loci where electrical stimulation evoked continuous decrease in IBAT temperature during the stimulation for 5 min were localized in and around the retrorubral field. Results suggest that tonic inhibitory mechanism on BAT thermogenesis exists in the homologous region of the hamster brain to the rat brain in the previous study.     

Effect of unilateral surgical denervation of brown adipose tissue on uncoupling protein mRNA level and cytochrom-c-oxidase activity in the Djungarian hamster

The bilateral lobe of interscapular brown adipose tissue of the Djungarian hamster was unilaterally denervated in order to study the role of the sympathetic innervation for maintenance and cold-induced increase of non-shivering thermogenesis. Denervation decreased the noradrenaline content of brown adipose tissue to less than 9% of the intact contralateral pad. This low noradrenaline level was maintained for 1–14 days after denervation. First, to study the role of the sympathetic innervation of brown adipose tissue in the maintenance of the high thermogenic capacity characteristic of the cold acclimated state, brown adipose tissue was denervated in hamsters either kept at thermoneutrality or acclimated to 5°C ambient temperature for 4 weeks. Cold-acclimated hamsters had elevated levels of uncoupling protein messenger ribonucleic acid (8.1-fold) and cytochrom-c oxidase-activity (3-fold). Denervation of brown adipose tissue decreased uncoupling protein-messenger ribonucleic acid level and cytochrom-c-oxidase-activity as compared to the intact pad in thermoneutral and in cold-acclimated hamsters. However, in cold-acclimated hamsters uncoupling protein-messenger ribonucleic acid level and cytochrom-c-oxidase-activity in denervated brown adipose tissue both were maintained on an elevated 6-fold higher levels as compared to thermoneutral controls. Second, to study the role of the sympathetic innervation of brown adipose tissue in the cold-induced increase in thermogenic capacity, hamsters were denervated prior to cold acclimation and responses were measured after 3 and 14 days of cold exposure. Uncoupling protein-messenger ribonucleic acid level and cytochrom-c-oxidase-activity of intact brown adipose tissue increased after 14 days cold acclimation. Denervation did not completely prevent a cold-induced 1.5-fold increase of cytochrom-c-oxidase-activity and a 3.2-fold increase of the uncoupling protein-messenger ribonucleic acid level in denervated brown adipose tissue after 14 days of cold acclimation. In conclusion, high levels of uncoupling protein-messenger ribonucleic acid and cytochrom-c-oxidase activity in brown adipose tissue of cold-acclimated hamsters can partially be maintained without intact sympathetic innervation, suggesting a considerable contribution of trophic factors not requiring sympathetic innervation for maintenance. The cold-induced increase of cytochrom-c-oxidase activity and expression of uncoupling protein-messenger ribonucleic acid largely depends upon sympathetic innervation of brown adipose tissue.Abbreviations ANOVA analysis of variance - BAT brown adipose tissue - COX cytochrom-c-oxidase - HPLC high performance liquid chromatography - mRNA messenger ribonucleie acid - NA noradrenaline - T _a ambient temperature - UCP uncoupling protein