Only UCP1 can mediate adaptive nonshivering thermogenesis in the cold.

Adaptive nonshivering thermogenesis may have profound effects on energy balance and is therefore therefore is a potential mechanism for counteracting the development of obesity. The molecular basis for adaptive nonshivering thermogenesis has remained a challenge that sparked acute interest with the identification of proteins (UCP2, UCP3, etc.) with high-sequence similarity to the original uncoupling protein-1 (UCP1), which is localized only in brown adipose tissue. Using UCP1-ablated mice, we examined whether any adaptive nonshivering thermogenesis could be recruited by acclimation to cold. Remarkably, by successive acclimation, the UCP1-ablated mice could be made to subsist for several weeks at 4C during which they had to constantly produce heat at four times their resting levels. Despite these extreme requirements for adaptive nonshivering thermogenesis, however, no substitution of shivering by any adaptive nonshivering thermogenic process occurred. Thus, although the existence of, for example, muscular mechanisms for adaptive nonshivering thermogenesis has recurrently been implied, we did not find any indication of such thermogenesis. Not even during prolonged and enhanced demand for extra heat production was any endogenous hormone or neurotransmitter able to recruit any UCP1-independent adaptive nonshivering thermogenic process in muscle or in any other organ, and no proteins other than UCP1-not even UCP2 or UCP3-therefore have the ability to mediate adaptive nonshivering thermogenesis in the cold.

Does cold acclimation induce nonshivering thermogenesis in juvenile birds? Experiments with Pekin ducklings and Japanese quail chicks.

The capability to produce heat in cold by nonshivering thermogenesis (NST) was studied in Pekin ducklings and Japanese quail chicks acclimated to cold for 3 weeks using indirect calorimetry (oxygen consumption) and electromyography from breast (M. pectoralis) and leg muscles (quails: M. gastrocnemius; ducklings: M. gastrocnemius, M. iliofibularis). Respiration of muscles in vitro was studied by measuring cytochrome c oxidase activity. In both species, cold acclimation induced clear morphometric and physiological changes, but no clear evidence of nonshivering thermogenesis. This was evident because increased shivering at least in one muscle coincided with increased oxygen consumption. In ducklings, however, amplitudes of shivering EMGs were low (<30 microV) in all muscles studied in both the control and cold-acclimated groups. Ducklings reacted to cold mainly by means of increasing body weight (1796 g in control, 2095 g in cold-acclimated) and circulatory changes. Acclimation did not change oxygen consumption either in vivo or in vitro. In quails, in addition to increased body weight (78.1 g control, 89.9 g cold-acclimated), improved insulation and metabolic adaptation to cold (increased respiration in vivo and in M. pectoralis in vitro) was also utilized. In Japanese quail chicks, 3 weeks of cold acclimation does not seem to induce NST, while in Pekin ducklings the existence of NST could not be totally excluded because of weak overall shivering activity.

Shivering and digestion-related thermogenesis in pigeons during dark phase.

The pigeon's main source of regulated heat production, shivering, is especially likely to be used for thermoregulation during the dark phase of the day when there is little heat from locomotor activity. However, food stored in the pigeon's crop is digested during the night, and digestion-related thermogenesis (DRT) will provide heat that should decrease the need for shivering to maintain body temperature (Tb). We investigated the conditions under which DRT alters the occurrence of nocturnal shivering thermogenesis in pigeons. In fasting experiments, in which DRT was minimal, variations in pectoral shivering were closely related to the kinetics of nocturnal Tb when the ambient temperature (Ta) was moderate (21 degrees C). In that case, shivering was low while Tb fell at the beginning of the night, moderate during the nocturnal plateau in Tb, and strong during the prelight increase in Tb. Similar kinetics of nocturnal Tb occurred when Ta = 28 degrees C, but shivering was negligible throughout the dark phase. In restricted feeding experiments, nocturnal DRT was varied by providing different amounts of food late in the light phase. When Ta = 21 degrees C, 11 degrees C, and 1 degrees C, nocturnal Tb and O2 consumption were directly related to the amount of food ingested. However, nocturnal shivering tended to decrease as the food load increased and was significantly reduced at the higher loads. Because nocturnal shivering did not become more efficient in producing heat as the size of the food load increased, we conclude that nocturnal DRT decreased the need for shivering thermogenesis.

Shivering thermogenesis in leg and breast muscles of galliform chicks and nestlings of the domestic pigeon.

We studied the ontogeny of shivering thermogenesis in breast and leg muscles of precocial galliforms (domestic fowl, grey partridge, and Japanese quail) and the altricial domestic pigeon using electromyography (EMG) and indirect calorimetry. Galliforms were able to increase heat production by shivering in leg muscles at the youngest age studied (1-2 d). Pectorals contributed to heat production from days 7-10 onward, but in the partridge and especially in the fowl, shivering by the pectorals was weaker than in the quail. In the pigeon, shivering began in pectorals and legs at 2 and 4 d of age, respectively, and pectorals had clearly the predominant role in thermogenesis. Despite the early beginning of electrical signs of shivering, significant thermogenesis did not appear in the pigeon before the age of 6 d. All galliforms shivered in bursts, like pigeons aged 2-4 d. From the age of 6 d onward, continuous shivering became predominant in the pigeon. In pectorals of 2-6-d-old pigeons, shivering did not increase linearly during decreasing ambient temperature, as in other muscles and species, but started abruptly, at full intensity. Furthermore, in 2-4-d-old pigeons, cooling induced movement activity in legs. The median frequency of shivering EMGs varied (1) with maturation of the muscle, (2) with size of the adult bird, and (3) between altricials and precocials.

Shivering thermogenesis in the pigeon: the effects of activity, diurnal factors, and feeding state.

Shivering (electromyographic activity of the pectoral muscle), oxygen consumption, and body temperature were measured from undisturbed pigeons for periods of several weeks, and segments from the midparts of each phase of the light-dark cycle were compared at various ambient temperatures and feeding regimes. Behavior was recorded with a video camera. None of the observed types of behavior (e.g., walking, preening, feeding, drinking, pecking, defecation) induced spurious electrical activity in the pectoral muscle. On the other hand, none of these behaviors directly inhibited ongoing shivering. There was no difference in the mean level of shivering between the light (L) and dark (D) phases of the day in any of the conditions, although body temperature was 2 degreesC higher during L. Measurements of integrated electromyogram (EMG) with high temporal resolution (28 samples/s) showed that, at 1 degreesC, shivering in the pectoral muscle was present for more than 98% of the time. Plots of oxygen consumption against root mean square EMG were obtained in each condition by a filtering procedure that excludes data points in which oxygen consumption is affected by motor activity. These plots showed that the increase in heat production induced by a unit increase in pectoral EMG was lower in D than in L and that it was further lowered by fasting. The amplitude spectra of raw EMG signals were similar in all conditions. Spectra of demodulated (rectified, low-pass filtered) EMG showed a distinct rhythmicity around 8 Hz at 21 degreesC that was further enhanced by fasting but absent at 1 degreesC. This suggests that the degree of synchronization and pattern of recruitment of motor units are specific for various temperatures and feeding regimes, and may partly explain the variable relation between heat production and muscle electrical activity. The results emphasize the advantages of long-term measurements for understanding the control of thermogenesis in birds.

Influence of cold shivering on fine motor control in the upper limb.

The aim of the investigation was to determine the effects of cold shivering on the accuracy of force output in distal, middle and proximal muscles of the upper limb. Test of hand grip strength, elbow flexion and shoulder flexion (each done at 10% maximal voluntary contraction for 15 s) were done under three conditions: (1) thermoneutral air (27 degrees C), a condition of thermal comfort; (2) cold air (10 degrees C), a condition eliciting an increase in tonic muscle activity; (3) and cold air (10 degrees C) with a cold drink (8 degrees C), a condition that causes visible shivering. The averaged (root mean square) electromyogram (AEMG) and mean power frequency (MPF) were measured from proximal, middle and distal arm muscles during the tests and compared. The control of force output was highly effective at thermoneutral condition for all motor tasks. During the cold air condition, all muscles were tonically active but there was no effect on accuracy of test performance. However, AEMG increased approximately 20% (P < 0.05) with respect to test performance in thermoneutral condition. During the cold air/cold drink condition, all muscles were shivering to a different extend. AEMG during test performance increased 30-150% in comparison to thermoneutral condition (P < 0.05). In this case, hand grip and elbow flexion were not adversely affected (these tests require middle and distal muscles) by cold shivering. However, the accuracy of performance of shoulder flexion was adversely affected. This is consistent with the fact that proximal muscles are more active during cold shivering.

Energy cost and thermoregulation of unrestrained rats during exercise in the cold.

The effect of cold ambient temperature on the energy cost and thermoregulation during exercise was studied in 18 male Sprague-Dawley rats accustomed to running for 2 weeks. For measuring T(b) and ensuring an unrestrained exercise performance, the rats were implanted with intraperitoneal telemetric radio transmitters. The rats ran at three submaximal exercise intensities (10, 15 and 20 m.min-1) at different T(a) values (22 degrees, 0 degree, -10 degrees and -20 degrees C) for 30 min. VO2 and Tb were continuously measured. During exercise at 0 degree C a 100% substitution of thermoregulatory heat production by exercise thermogenesis was observed. At T(a) values below 0 degree C the physical strain of exercise increased considerably: Vo2 increased 34-61% at -10 degrees C and 51% at -20 degrees C compared to the corresponding exercise intensity at T(a) 22 degrees C. A partial substitution of thermoregulatory heat production by exercise thermogenesis was observed in the cold. At -10 degrees C the cost for thermoregulation during exercise was 28% and at -20 degrees C 31% lower than at rest at the corresponding T(a) values. The rat could not maintain normal Tb during exercise below 0 degrees C. The T(b) dropped 1.5-1.8 degrees C at -10 degrees C and 2.5 degrees C at -20 degrees C. It is concluded that exercising below 0 degree C at submaximal levels (below 50% of VO2max) partially substitutes for thermoregulatory costs but is thermally unfavorable for the small-sized rat.

Seasonal patterns in the physiology of the European brown bear (Ursus arctos arctos) in Finland.

The physiological indicators such as body temperature, blood chemistry and hematology of seven European brown bears (Ursus arctos arctos) were used in the present study. They were kept in either the Zoological Garden of University of Oulu (65 degrees N, 25 degrees 24'E) or the Ranua Zoological Garden approx. 150 km NE of Oulu. Transmitters with a temperature-dependent pulse rate were implanted subcutaneously or into the abdominal cavity under anesthesia. Our data indicate that the body temperature of the bear decreases during the winter sleep to 4-5 degrees C below the normal level (37.0-37.5 degrees C). The lowest values, 33.1-33.3 degrees C, were measured several times in midwinter. Hematocrit, hemoglobin and erythrocyte counts seem to be higher, and the leucocyte count lower during the denning period than in the awake bear. Plasma N-wastes were lower during the winter sleep than before or after it. The analysed blood parameters showed that plasma catecholamines and thyroid hormones decreased in the fall.

Nocturnal hypothermia in fasting Japanese quail: the effect of ambient temperature.

Japanese quail were equipped with intraperitoneal transmitters for telemetric measurement of body temperature (Tb) and activity. Food deprivation at + 24 degrees C for four days induced a well-defined nocturnal hypothermic response. The normal day-night difference (about 1 degrees C) in Tb increased 1 degrees C per day, reaching 5 degrees C on the fourth night of fast. Nocturnal motor activity decreased during the fast, while daytime activity first increased and then returned to the original level by the end of the fasting period. A strong correlation between Tb and motor activity was found during scotophase (r = .91) but not during photophase (r = .02). At + 4 degrees C, where food was deprived for two days, the level of hypothermia was not different from that at + 24 degrees C, but birds fasting in the cold reached the hypothermic level more rapidly. In control birds, the decrease in Tb at the beginning of scotophase was independent of ambient temperature. In the combined data, the level of hypothermia correlated strongly with body mass loss (r = .90), which shows that quail can directly or indirectly sense the amount of body energy reserves. This is the first report of hypothermia in a fasting gallinaceous bird. The consistent level of hypothermia at varying ambient temperatures suggests that either nonenergetic costs or phylogenetic constraints prevent deeper hypothermia in cold. Accordingly, the regulation of hypothermia cannot be explained by using only energetic arguments.

Is the "mammalian" brown fat-specific mitochondrial uncoupling protein present in adipose tissues of birds?

1. Mitochondria were isolated from the furcular, subcutaneous, abdominal, nape and lateral adipose tissue depots of five species of bird (pheasant, Japanese quail, pigeon, house sparrow and great tit) acclimatized to the Northern winter. 2. Mitochondrial proteins were separated by SDS-polyacrylamide gel electrophoresis, blotted onto nitrocellulose membranes, and probed for the presence of the 32,000-33,000 Mr uncoupling protein characteristic of "mammalian" brown adipose tissue, using an anti-(ground squirrel uncoupling protein)serum. 3. Immunoreactivity consistent with uncoupling protein was not detected in mitochondria from any of the avian adipose tissues. Immunoreactivity was, however, evident in mitochondria from perirenal or interscapular adipose tissue from a range of mammals--rats, mice, golden hamsters, Orkney voles, wood mice, pipistrelle bats, wood lemmings, and newborn lambs, cattle, reindeer and red deer. 4. These results provide biochemical evidence that "mammalian-like" thermogenic brown adipose tissue is absent from avian species; adipose tissues in birds appear to be functionally "white".

Respiratory modulation of shivering intensity in the pigeon.

Respiration and shivering were measured in unanaesthetized, cold-exposed pigeons using pneumotachography and electromyography, respectively. The instantaneous intensity of shivering in the pectoral muscle varied in phase with respiration. Power spectral analysis showed that the main frequency components of respiration and demodulated EMG coincided exactly. The intensity of shivering was highest during end-expiration and lowest at end-inspiration. This was confirmed by cross-correlation analysis of respiration and demodulated EMG. The absolute level of modulation remained constant (c. 10 microV peak-to-peak) despite changes in the general intensity of shivering. On the other hand, the relative depth of modulation was highest during incipient shivering. These facts indicate that only a part of the motor units recruited for shivering is susceptible to respiratory modulation and that this part is first recruited during incipient shivering. Inhalation of 5% CO2 did not affect the interaction between respiration and shivering although respiration frequency varied from 25 to 60 min-1. Thus, pulmonary chemoreceptors do not mediate this effect. It is suggested that the interaction between respiration and shivering occurs directly in the CNS. The question whether the interaction is adaptive for the animal or merely reflects a common evolutionary history of the underlying neural circuits is discussed.

The relationship of muscle electrical activity, tremor and heat production to shivering thermogenesis in Japanese quail.

We measured the electrical activity and the tremor of the pectoral muscle and total body heat production in control and cold-acclimated Japanese quail at +26 degrees C, +12 degrees C and +2 degrees C before and after 3 weeks of acclimation, using electromyography, accelerometer recordings, and indirect calorimetry. Japanese quail shiver in 0.2- to 3-s bursts that occur in groups. An increase in both the frequency and the duration of bursts and burst groups contributes to the increase in heat production by shivering at low temperatures. A compilation of shivering patterns in birds is given and its implications for the neural control and phylogeny of shivering are discussed. A rather non-specific increase in electromyographic (EMG) activity and heat production was observed after cold acclimation at all experimental temperatures, although many of the normal signs of cold acclimation (e.g. decrease in gonad mass, increase in heart mass and serum triiodothyronine) were seen. The increase in muscle electrical activity was greater than the increase in oxygen uptake, which resulted in a lower VO2/EMG ratio. The amplitude distribution of muscle electrical activity remained normal, but a shift towards higher frequencies occurred in the EMG spectra of cold-acclimated birds. Despite the increase in muscle electrical activity, power spectra of accelerometer recordings indicated that the amplitude of the muscle tremor was lower in cold-acclimated birds. The increase in the high-frequency components of the EMG indicates that decreased synchronization of motor unit firing may account for the lower tremor amplitude. We suggest that this change is adaptive because it reduces heat loss and/or because more fatigue-resistant motor units are recruited. These results show that temperature acclimation modifies the neural control of shivering in skeletal muscle.

Response to cold in the blue fox and raccoon dog as evaluated by metabolism, heart rate and muscular shivering: a re-evaluation.

Oxygen consumption (ml kg-0.75/min) in relation to ambient air temperature at or below the lower critical temperature (Tlc) of the winter-furred raccoon dog (+10 degrees C) and the blue fox (-6 degrees C) is described by the equations y = 14.8-0.28x and y = 7.5-0.20x, respectively. Muscular shivering activity (integrated EMG) of both species increased below thermoneutrality parallel with increasing oxygen uptake and heart rate. Seasonal changes in measured metabolic parameters were evident for both species. The results suggest that the overall body insulation or the metabolic response to cold are not essentially worse in the raccoon dog as compared with the blue fox. It is concluded that earlier speculations of surprisingly wide thermoneutral zone and very low Tlc of the Arctic fox are not evident for the blue fox.

Related and unrelated changes in response to exercise and cold in rats: a reevaluation.

Groups of rats were subjected to various treatments: continuous exposure to cold (5 degrees C); exercise by running; intermittent cold exposure, -20 degrees C daily for 60 min; and in some experiments combined influence of cold acclimation and exercise for at least 6 wk. The resulting adaptive changes can be grouped in three different categories. Cold-specific changes included increased food intake, an increase in both mass and metabolic activity of brown adipose tissue leading to an increased capacity for nonshivering thermogenesis, and maintenance of the stores of ascorbic acid and muscle glycogen during cold exposure. These changes were associated with an improved resistance to cold with which the rats were able to maintain their body temperature in both cold air and water were typical of rats previously exposed to cold. Training-specific changes typically included increased activities of aerobic muscle enzymes and decreased activity of lactate dehydrogenase and a higher O2 uptake and shivering activity during cold exposure as compared with sedentary control rats. These changes were observed for trained rats only and were not associated with an improved resistance to cold. Other adaptive changes were found, to a variable extent, for all treated rat groups. These included cardiac hypertrophy, reduced urinary catecholamine excretion during and after stress situations, increased tail skin temperature response to isoproterenol, and a higher tail skin temperature during exposure to cold. There were no systematic differences between groups in changes of blood glucose, glycerol, or lactate concentrations during cold exposure.(ABSTRACT TRUNCATED AT 250 WORDS)

Thermal and electromyographic correlates of shivering thermogenesis in the pigeon.

1. Electromyographic, thermal and metabolic measurements were made on pigeons subjected to stepwise ambient cooling. Subsequent signal analysis of the EMG-recordings from the pectoral muscle showed that the mean rectified value (Umrv) is the most reliable EMG-based indicator of metabolic heat production (M). 2. The muscle-abdomen temperature difference was linearly related to M, and correlated strongly with Umrv. 3. The long-term and short-term fluctuations of shivering intensity (Umrv) were highly synchronous in the two main muscles, m. pectoralis and m. supracoracoideus. 4. The EMG-signal approached a Gaussian process at high shivering intensities. Evaluation of the changes in EMG median frequency suggested a size-dependent recruitment of motor units for shivering. 5. It is pointed out that Umrv and the integrated value of EMG are interconvertible and that Umrv should be preferred, because it has more physiological relevance.

Effects of blood pressure manipulations on shivering thermogenesis in the pigeon.

The effects of blood pressure (BP) manipulations on shivering thermogenesis were studied in conscious pigeons. A rise in BP induced by noradrenaline (NA) or equipotent doses of angiotensin II (Ang II) effectively suppressed shivering at +12 degree C and partly abolished the cold-induced vasoconstriction in the feet. The inhibition commenced when the rise in BP reached +40 mmHg, and a fall in body temperature followed these responses. Comparison of the trajectories in the BP-shivering plane revealed that the inhibition of shivering by Ang II could be completely explained by changes in BP, whereas NA had also another, more prolonged inhibitory action independent of baroreceptor activity. A similar dose-dependence for effects on BP and shivering could be established with both drugs. An acute reduction of BP by sodium nitroprusside had also a very potent inhibitory effect on shivering. In hypotensive pigeons elevation of BP with NA initially enhanced shivering, but when BP was raised beyond normal levels shivering was again suppressed. We conclude that both a rise and a fall in BP can inhibit shivering in the pigeon, and that normal levels of BP facilitate shivering. NA inhibits shivering by more than one mechanism, but the initial effect is mediated through a baroreflexive action. The interactions of thermoregulatory and cardiovascular mechanisms suggest an integrated control of body temperature and circulation, which should be considered in experimental approaches to these homeostatic systems.

Differential impairment of thermogenesis in the pigeon after chemical sympathectomy.

A dose-controlled chemical sympathectomy with 6-hydroxydopamine (6-OHDA) did not disrupt thermostasis in the pigeon at +38 degrees C. At +6 degrees C, thermogenesis was impaired, but the lower body temperature and oxygen consumption were stable and vasoconstriction was normal. The stability may partly be explained by a massive release of adrenaline from the adrenals (50% in 20 min). Despite a deficit in heat production both after sympathectomy and after acute 6-OHDA, no change in muscle electrical activity was observed. Plasma free fatty acid (FFA) concentration was significantly elevated after sympathectomy, but no changes occurred in blood glucose or plasma lactate levels. The results indicate a major compensatory role for the adrenals in avian thermogenesis. They also suggest a sympathetically mediated auxiliary thermogenic mechanism independent of muscle electrical activity and coupled to FFA metabolism.