Shivering and nonshivering thermogenesis in exercised cold-deacclimated rats

Norepinephrine (NE)-induced increase in oxygen consumption (VO2) and colonic temperature (Tc) was greater in cold-acclimated rats housed at 4 degrees C for 4 weeks (CA) than warm-acclimated controls housed at 24 degrees C for 4 weeks (WA). On the other hand, shivering activity measured at 4 degrees C was less in CA than in WA, while propranolol administration eliminated the difference between these two groups by enhancing shivering in CA. Wet weight and protein content of interscapular brown adipose tissue (IBAT) were greater in CA than in WA. Following cold acclimation, CA were deacclimated at 24 degrees C for 5 weeks. During deacclimation, half of this latter group were forced to run (15 m.min-1 for 1 h) every day (CD-T) while the remaining rats remained sedentary (CD-S). Shivering activity assessed at 4 degrees C 4 weeks after commencing cold deacclimation was significantly less in CD-T than in CD-S and the difference disappeared following propranolol injection. VO2 and Tc responses to NE injection measured 1, 2 and 5 weeks after commencing cold deacclimation did not differ between CD-S and CD-T. Although IBAT weight was lighter in CD-T than in CD-S, its total protein content was not different between the latter two groups of rats. These results suggest that a greater degree of NE-independent nonshivering thermogenesis (NST) is retained in rats that are exercised during the process of deacclimation as compared with animals that are sedentary. This difference in NST would not seem to be directly related to BAT thermogenic capacity.     

Effect of different environmental temperatures on the serotonin concentration and turnover in the brain stem of a hibernator.

The serotonin (5-HT) and 5-hydroxyindoleacttic acid (5-HIAA) levels and 5-HT turnover were studies in the brain stem of warm- (+30 degrees C) and cold- (+6 degrees C) acclimated golden hamsters, exposed for 3 hours to temperatures of +6 degrees C, +30 degrees C and +37 degrees C, respectively. In war-acclimated hamsters kept under conditions the 5-HT level in the brain did not change significantly during the year. The 5-HIAA level was slightly higher in the winter. The 5-HT turnover varied within limits of 0.071 to 0.180 mug/g/hour-1. Three hours' exposure of warm-acclimated golden hamsters to cold (6 degrees C) increased the concentrations of 5-HT and 5-HIAA and the 5-HT turnover in the brain. After long-term adaptation to cold (6 degrees C) the 5-HT level, and the 5-HT turnover returned to the original level. Three hours' exposure of golden hamsters to higher environmental temperatures (warm-acclimated individuals to 37 degrees C and cold-acclimated individuals to 30 degrees C) also increased the 5-HT turnover. The concentrations of 5-HT and 5-HIAA increased in cold-acclimated golden hamsters exposed to 30 degrees C and was not changed in warm-acclimated ones, exposed to 37 degrees C. Although the elevated temperatures induce greater changes in serotonin metabolism than lowered temperatures, the serotonin pathways in the brain do not seem to be affected by short-term temperature changes specifically. The findings are rather indicative that changes in 5-HT turnover may be the primary reaction to stressful conditions.     

Cold acclimation increases basal heart rate but decreases its thermal tolerance in rainbow trout (Oncorhynchus mykiss)

Rainbow trout (Oncorhynchus mykiss, Walbaum) were acclimated to 4 degrees C and 17 degrees C for more than 4 weeks and heart rate was determined in the absence and presence of adrenaline to see how thermal adaptation influences basal heart rate and its beta-adrenergic control in a eurythermal fish species. The basal heart rate in vitro was higher in cold-acclimated than warm-acclimated rainbow trout at temperatures below 17 degrees C. On the other hand, adaptation to cold decreased thermal tolerance of heart rate so that the maximal heart rates were achieved at 17 degrees C (75 +/- 4 bpm) and 24 degrees C (88 +/- 2 bpm) in cold-acclimated and warm-acclimated trout, respectively. Beta-adrenergic response of the heart was enhanced by cold-adaptation, since adrenaline (100 nmol l(-1)) caused stronger stimulation of heart rate in cold-acclimated (29 +/- 14%) than in warm-acclimated fish (10 +/- 1%; P = 0.03). Furthermore, adrenaline strongly opposed the temperature-dependent deterioration of force production in cold-acclimated trout but not in warm-acclimated trout. The results indicate that adaptation to cold increases basal heart rate but decreases its thermal tolerance in rainbow trout. Cold acclimation up-regulates the beta-adrenergic system, and beta-adrenoceptor activation seems to provide cardioprotection against high temperatures in the cold-adapted rainbow trout.     

Thermoregulatory responses of dystrophic hamsters to changes in ambient temperatures

The ability of dystrophic hamsters to maintain their body temperature despite abnormal muscle and brown adipose tissue, two organs involved in thermoregulation, was evaluated. Dystrophic hamsters (CHF 146) between the ages of 30 and 160 days kept at 21 degrees C had core (rectal) temperatures (TR) that were 0.5-1.5 degrees C lower than Golden Syrian controls. The reduced core temperatures of dystrophic hamsters were unlikely the result of an incapacity to generate heat since the dystrophic hamsters were able to maintain their TRs during 3 h of acute cold stress (4 degrees C) and to adapt to prolonged cold exposure. However, TRs of cold-acclimated dystrophic hamsters were still 1 degree C below TRs of cold-acclimated control animals. By contrast, increasing the ambient temperature raised TRs of both normal and dystrophic hamsters. When kept at 32 degrees C overnight, the TRs of dystrophic hamsters remained significantly below those of control animals. When heat-exposed dystrophic hamsters were returned to 21 degrees C, their TRs returned to values significantly lower than those of control hamsters. Thus, dystrophic hamsters showed a capacity to thermoregulate, like control hamsters, but appeared to do so at a lower temperature. The reduced core temperatures of dystrophic hamsters kept at 21 degrees C cannot be explained by a reduction in metabolic activity since newborns and 30- and 140-day-old dystrophic hamsters had rates of oxygen consumption (VO2) and carbon dioxide production (VCO2) that were similar to those of controls. These results suggest that the thermoregulatory set point may be altered in dystrophic hamsters.     

Fatty acid and sterol synthesis by hepatocytes of thermally acclimated rainbow trout (Salmo gairdneri).

Incorporation of tritium from tritiated water into lipid fractions was measured in isolated hepatocytes from rainbow trout (Salmo gairdneri) acclimated to 5 degrees C and 20 degrees C. Hepatocytes from cold-acclimated trout exhibited significantly higher rates of tritium incorporation into both fatty acid and sterol fractions at assay temperatures of 15 degrees C and 20 degrees C than did hepatocytes from warm-acclimated trout. Tritium incorporation into the fatty acid fraction was nearly temperature independent in hepatocytes from warm-acclimated trout (Q10 = 1.39) but markedly temperature dependent (Q10 = 2.63) in hepatocytes from cold-acclimated trout; in contrast, rates of sterol synthesis were more temperature dependent in warm-acclimated trout. At 5 degrees C, fatty acid lipogenesis comprised a significantly greater percentage of the total tritium incorporation in hepatocytes from warm-acclimated trout and the percentage of total lipogenesis attributable to fatty acids decreased significantly in warm-acclimated trout as the assay temperature increased; the opposite trends were observed in cold-acclimated trout.     

Effects of consumption of juniper (Juniperus monosperma) on cost of thermoregulation in the woodrats Neotoma albigula and Neotoma stephensi at different acclimation temperatures

A study was done to test whether toxic plants that occur naturally in the diet affect thermoregulation in mammalian herbivores. The woodrats Neotoma albigula and Neotoma stephensi both consume juniper (Juniperus monosperma), a plant with high levels of toxic compounds. Body temperature (Tb), basal metabolic rate (BMR), and the minimum cost of thermogenesis (Cmin) were measured for both species on control and juniper diets following warm (25 degrees C) and cold (18 degrees C) acclimation. In N. albigula, diet had no uniform effect on Tb, BMR, or Cmin, but dietxacclimation-temperature interaction effects on Tb and Cmin were highly significant (P<0.005). For thermoregulation at 15 degrees C, juniper consumption increased the metabolic cost for warm-acclimated N. albigula by 50% but decreased the metabolic cost in cold-acclimated N. albigula by 24%. In N. stephensi, diet significantly affected Tb and Cmin (P<0.05), but there were no significant dietxacclimation-temperature interaction effects. For thermoregulation at 15 degrees C, juniper consumption increased the metabolic cost for warm-acclimated N. stephensi by 33% but had no significant effect on metabolic cost in cold-acclimated N. stephensi.     

Effects of exposure temperature on brown adipose tissue uncoupling protein mRNA levels

The effect of temperature on the amount of uncoupling protein mRNA in rat brown adipose tissue was examined after 1 and 14 days of exposure to cold. The relative amounts after 1 day, compared with rats kept at a thermoneutral temperature of 28 degrees C, were 3.2 at 19 degrees C, 3.3 at 11 degrees C, and 2.1 at 3 degrees C. This suggests that in warm-acclimated rats, a maximal response to a cold stimulus in brown adipose tissue is reached by 19 degrees C. In contrast to these results, the relative amounts of uncoupling protein mRNA after 14 days of cold exposure, compared with rats left at 28 degrees C, were 1.2 at 19 degrees C, 1.9 at 11 degrees C, and 2.1 at 3 degrees C. Since it is known that the amount of uncoupling protein in cold-acclimated rats increases continuously with decrease in temperature, the amount of protein reflects the mRNA levels during later times but not the initial time of exposure to cold.     

Temperature acclimation modifies sinoatrial pacemaker mechanism of the rainbow trout heart

The hypothesis of pacemaker level origin of thermal compensation in heart rate was tested by recording action potentials (AP) in intact sinoatrial tissue and enzymatically isolated pacemaker cells of rainbow trout acclimated at 4 degrees C (cold) and 18 degrees C (warm). With electrophysiological recordings, the primary pacemaker was located at the base of the sinoatrial valve, where a morphologically distinct ring of tissue comprising myocytes and neural elements was found by histological examination. Intrinsic beating rate of this pacemaker was higher in cold-acclimated (46 +/- 6 APs/min) than warm-acclimated trout (38 +/- 3 APs/min; P < 0.05), and a similar difference was seen in beating rate of isolated pacemaker cells (44 +/- 6 vs. 38 +/- 6 APs/min; P < 0.05), supporting the hypothesis that thermal acclimation modifies the intrinsic pacemaker mechanism of fish heart. Inhibition of sarcoplasmic reticulum (SR) with 10 microM ryanodine and 1 microM thapsigargin did not affect heart rate in either warm- or cold-acclimated trout at 11 degrees C but reduced heart rate in warm-acclimated trout from 74 +/- 2 to 42 +/- 6 APs/min (P < 0.05) at 18 degrees C. At 11 degrees C, a half-maximal blockade of the delayed rectifier K+ current (I(Kr)) with 0.1 microM E-4031 reduced heart rate more in warm-acclimated (from 45 +/- 1 to 24 +/- 5 APs/min) than cold-acclimated trout (56 +/- 3 vs. 48 +/- 2 APs/min), whereas I(Kr) density was higher and AP duration less in cold-acclimated trout (P > 0.05). Collectively, these findings suggest that a cold-induced increase in AP discharge frequency is at least partly due to higher density of the I(Kr) in the cold-acclimated trout, whereas contribution of SR Ca2+ release to thermal compensation of heart rate is negligible.     

Rapid cold-induced changes of membrane order and delta 9-desaturase activity in endoplasmic reticulum of carp liver: a time-course study of thermal acclimation

The membrane order of liver endoplasmic reticulum (ER) membranes of 10 degrees C- and 30 degrees C-acclimated carp has been compared using the fluorescence polarization technique with DPH as probe. Membranes from cold-acclimated fish displayed lower polarizations than corresponding membranes from warm-acclimated fish, the difference compensating for 34-50% of the direct effects of temperature upon polarization. The changes in delta 9-desaturase activity and fluorescence polarization of DPH in ER membranes have been monitored as a function of time during cold acclimation of 30 degrees C-acclimated carp. Cooling was achieved in three stages over 48 h. Desaturase activity in both rough and smooth ER showed a rapid increase in activity for the first three days followed by a decline on day 4 and a second increase up to day 10. Polarization of DPH (measured at 10 degrees C) was rapidly reduced on cooling with no further change after day 4. The halftime for change in polarization and for the first desaturase induction were both approx. 2 days although large changes in polarization were evident within 24 h after the onset of cooling. During the cooling phases the daily changes in DPH polarization were quantitatively related to increments in desaturase capacity. The second desaturase induction had no effect upon membrane structure, at least as indicated by the polarization technique.     

Aerial and aquatic oxygen uptake by freely-diving snapping turtles (Chelydra serpentina)

Summary Aerial oxygen consumption of unrestrained, freely-diving warm-and cold-acclimated snapping turtles, Chelydra serpentina, was measured at 10, 20, and 30°C. Also, simultaneous determinations of aerial and aquatic oxygen uptake by voluntarilydiving animals were made at 4 and 20°C. The standard rates of aerial oxygen consumption are equivalent in cold-and warm-acclimated animals in water and in cold-acclimated ones in air; these rates are all lower than those of warm-acclimated animals in air. Thus either cold acclimation or voluntary submergence reduces the standard metabolic rate of snapping turtles but the effects are not additive. Aquatic oxygen uptake during voluntary submergence is more important at low than at moderate temperatures and probably contributes significantly to gas exchange in these animals as they overwinter beneath the ice of ponds and streams.     

The effect of acclimation on mating frequency and mating competitiveness in the Queensland fruit fly, Dacus tryoni, in optimal and cool mating regimes

Mating frequency in groups of Dacus tryoni which had been either warm-acclimated or cold-acclimated were compared in temperature regimes ranging from just above mating-threshold to optimal. Cold-acclimation appeared to suppress initial mating ability of mature insects of both sexes to an extent which depended upon the acclimation regime used. The most favourable cold-acclimation regime produced flies which in certain circumstances were able to mate at an initial frequency similar to that of warm-acclimated flies. In no mating regime was initial mating significantly more frequent in any cold-acclimated group than it was in any warm-acclimated group. In most cases warm-acclimated flies in a given regime mated at high frequency for 1–2 days, whereas the cold-acclimated flies mated at low frequency for a greater number of days. In all cases, cold-acclimated flies accumulated a similar or significantly lower total number of matings than warm-acclimated groups. In experiments in which both warm-acclimated and cold-acclimated males competed for cold-acclimated females, the warm-acclimated males always out-competed the cold-acclimated males in two mild (near optimal) regimes. In a relatively harsh (near torpor threshold) regime, there was no significant difference in the competitive abilities of cold-acclimated and warm-acclimated males. The relevance of these results to possible acclimation procedures used in control campaigns involving release of sterile males is discussed.     

ATP-sensitive K+ channels in cardiac muscle from cold-acclimated goldfish: characterization and altered response to ATP

ATP-sensitive potassium channels (K(ATP)) play an important, if incompletely defined, role in myocardial function in mammals. With the discovery that K(ATP) channels are also present at high densities in the hearts of vertebrate ectotherms, speculation arises as to their function during periods of cold-acclimation and depressed ATP synthesis. We used single-channel and intracellular recording techniques to examine the possibility that channel activity would be altered in cardiac muscle from goldfish (Carassius auratus) acclimated at 7+/-1 degrees C relative to control (21+/-1 degrees C). As previously observed in mammals, K(ATP) channels in isolated ventricular myocytes were inwardly rectified with slope conductances of 63 pS. However, channel mean open-time and overall open-state probability (Po) were significantly increased in cells from the cold-acclimated animals. In addition, K(ATP) channels in cells from fish acclimated at 7 degrees were nearly insensitive to the inhibitory effects of 2 mM ATP, whether studied at 7 or at 21 degrees C. Transmembrane action potential duration (APD) in hearts of cold-acclimated fish studied at 21 degrees was significantly shorter than that observed in hearts of warm-acclimated fish at the same temperature; this difference was eliminated by the K(ATP) channel antagonist glibenclamide (5 microM). These data suggest that K(ATP) channels in the hearts of cold-acclimated animals are more active and less sensitive to ATP-inhibition than those in warm-acclimated fish, possibly reflecting a functional adaptation to promote tolerance of low temperatures in this species.     

Effects of adenosine on the contractility of normoxic rainbow trout heart

Temperature strongly affects oxygen solubility in water, oxygen convection in the blood and locomotor activity of the fish. Since oxygen supply and demand are temperature dependent, it was hypothesized that the purinergic control of the heart, one of the most important mediators in oxygen-limited conditions, might also show temperature dependence. Therefore, the present study examines the effects of adenosine (Ado), a purinergic agonist, on the contractile and electrical activity of the thermally acclimated trout ( Oncorhynchus mykiss Walbaum) heart. The fish were acclimated to either 4 degrees C or 17 degrees C and the experiments were conducted at the acclimation temperatures of the animals. In spontaneously beating hearts, Ado had a negative chronotropic and a positive inotropic effect in warm-acclimated rainbow trout while no response was detected in cold-acclimated trout. In paced atrial and ventricular preparations, Ado had a negative inotropic effect in both warm- and cold-acclimated fish, and the response was strongest in the atria of warm-acclimated trout. Ado shortened the duration of contraction 12-14% in atrial preparations but had no effect in ventricular muscle. Ado (10(-4) mol l(-1)) increased the density of the inwardly rectifying K(+) current from -3.5+/-0.6 pA pF(-1) to -8.4+/-1.4 pA pF(-1) (at -120 mV) in atrial myocytes of warm-acclimated trout but was without effect in atrial myocytes of cold-acclimated trout (-2.4+/-0.8 pA pF(-1) vs. -2.1+/-0.9 pA pF(-1)). Ado had no effect on K(+) currents of ventricular cells in either acclimation group. These results indicate that the effects of Ado on cardiac contractility and electrical activity are stronger in warm-acclimated than in cold-acclimated trout when measured at the physiological body temperatures of the fish. The balance between oxygen demand and supply of the heart might be better in the cold where more environmental oxygen is available and the power of the muscles is weaker thereby reducing the need for the purinergic control of the heart. Temperature-dependence of Ado response in the trout heart warrants that temperature should be taken into consideration when the purinergic system of the ectotherms is studied.     

Structural studies of natural actomyosin from thermally acclimated frogs.

Natural actomyosin was isolated from skeletal muscle of frogs (Rana catesbeiana) acclimated at 25 degrees C and 5 degrees C. It was found that preparations isolated from warm-acclimated frogs may display considerable degradation of myosin heavy chains as compared with preparations isolated from cold-acclimated frogs. However, degradation may be minimized by inclusion of protease inhibitors during purification, indicating enhanced protease activity in preparations of natural actomyosin from warm-acclimated frogs. When purified in the presence of protease inhibitors, natural actomyosin from both warm-acclimated and cold-acclimated frogs exhibits comparable subunit composition of SDS-gel electrophoresis. The overall gel pattern is similar to that obtained from rabbit natural actomyosin except that in the frog, troponin-T and troponin-C appear to co-migrate with tropomyosin and myosin light chain 2, respectively.     

Characterization of gill Na/K-ATPase activity and ouabain binding in Antarctic and New Zealand nototheniid fishes

The effects of thermal acclimation in two Nototheniid species, the stenothermal Antarctic Trematomous bernacchii and the eurythermal New Zealand Notothenia angustata, were investigated. Serum osmolality, gill Na/K-ATPase activity, sodium pump density and ouabain affinity were determined. Both fish were acclimated at their upper and lower viable thermal temperatures. Warm acclimation (+4 degrees C) of the T. bernacchii significantly decreased their serum osmolality from 550 to 450 mOsm/kg compared to cold-acclimation (-1.5 degrees C) and this was accompanied by a two-fold increase in gill Na/K-ATPase activity. Warm-acclimation (+14 degrees C) of N. angustata did not significantly change their serum osmolality from 330 mOsm/kg or gill Na/K-ATPase activity compared to the cold-acclimated (+4 degrees C) N. angustata. Using [(3)H]ouabain binding techniques, the B(max) and K(d) values of gill Na/K-ATPase enzymes were determined. No difference in the B(max) or K(d) of the warm-acclimated T. bernacchii accounted for the increase in Na/K-ATPase activity. We conclude that the change in gill Na/K-ATPase activity in the warm-acclimated T. bernacchii is not mediated by an increase in the number of enzyme sites and is not reflected in a change in ouabain affinity for Na/K-ATPase.     

Plasma free fatty acid levels during cold-induced and noradrenaline-induced nonshivering thermogenesis in the Djungarian hamster

Summary In Djungarian hamsters the cold-induced thermoregulatory heat production was preceeded and accompanied by an increase in the plasma level of free fatty acids. In warm-acclimated hamsters this increase was found more pronounced (0.85 to 1.48 mM) than in cold-acclimated hamsters (0.64 to 0.88 mM). Noradrenaline-induced thermogenesis at thermoneutrality provoked a similar increase in the free fatty acid level. Inhibition of nonshivering thermogenesis during cold exposure by propranolol abolished the increase in free fatty acids completely. The surgical removal of brown adipose tissue proportionately reduced the increase in free fatty acids. This indicates that the rise in plasma free fatty acids is functionally related to nonshivering thermogenesis and originates from brown adipose tissue.     

Some aspects of acclimation to low temperatures in the grain weevils Sitophilus oryzae (L.) and S. granarius (L.)

The ability of Sitophilus oryzae (L.) and S. granarius (L.) to acclimate when transferred from 27°C, a near optimal temperature, to 15° C, a marginal temperature, was examined in terms of chill-coma temperature, dispersal, oxygen consumption and rate of oviposition. The chill-coma temperature of S. oryzae was higher and the increment of acclimation less than that of S. granarius. Acclimation, although well advanced after 2 weeks after transfer, took more than 4 weeks to complete. Re-acclimation after the reciprocal transfer took only 2 weeks. The dispersal of S. oryzae held at 15 and 27° C prior to testing differed when the weevils were released in wheat at 27°C but not when released at 15°C. Cold-acclimated S. granarius dispersed less than warm-acclimated weevils when released in wheat at both 15 and 27°C. The activity oxygen consumption of cold-acclimated weevils was less than that of warm-acclimated weevils. Inverse-shifts of acutely determined R-T relationships appeared more significant than changes in temperature sensitivity. Acclimation to 15° C took 8–10 days and re-acclimation to 27°C about 6–8 days. S. oryzae consumed less oxygen per unit of weight than S. granarius but was more temperature sensitive. Resting oxygen consumption, which was less temperature sensitive than activity consumption, also exhibited inverse acclimation in both species. S. granarius showed evidence of inverse acclimation of oviposition rate after acclimation at 15°C.     

The Effects of Cold Acclimation upon the Oxygen Consumption of Two Species of Free-Living Nematodes

Two species of free-living nematodes, Panagrellus redivivus and Turbatrix aceti, were cultured axenically at control (20 C) and cold (10 C) temperatures. Oxygen consumption of worms from each population was measured manometrically on days 2 through 8 after exposure to these temperatures. In both species, the slope of the oxygen consumption curve for the controls was greater than that of the worms exposed to the cold on day 2. The slope of the curve of the cold-exposed worms gradually increased until day 7. At this time, the slope of the oxygen consumption curve from the cold-exposed worms exceeded or equaled that of the controls. This is taken as an indication of the onset of the cold-acclimated state in both species of worms by day 7.     

Deer mouse aerobic performance across altitudes: effects of developmental history and temperature acclimation

Aerobic physiology at high altitudes has been studied in many animals. Prior work on laboratory-bred deer mice (a species with a wide altitudinal range) showed depression of aerobic capacity at high altitude, even after acclimation. However, wild deer mice show no reduction in thermogenic performance at high altitude, and performance limits seem to be due to physiological and anatomical adjustments to environmental temperature and not to oxygen availability. We asked whether across-altitude performance differences exist in deer mice after accounting for temperature acclimation (approximately 5 degrees and 20 degrees -25 degrees C) and prenatal and neonatal development altitude (340 vs. 3,800 m). We measured maximal thermogenic oxygen consumption (VO2sum) in cold exposure and ran mice on a treadmill to elicit maximal exercise oxygen consumption (VO2max). We found a 10% reduction in VO2max at 3,800 m compared with that at 340 m; thus, the mice were able to compensate for most of the 37% reduction in oxygen availability at the higher altitude. Development altitude did not affect VO2max. There was no effect of test altitude or development altitude on VO2sum in warm-acclimated animals, but both test and development altitude strongly affected VO2sum in cold-acclimated mice, and compensation for hypoxia at 3,800 m was considerably less than that for exercise.