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Age comparisons of body temperature and cold tolerance among different strains of Mus musculus.

The age-related declines in colonic temperature (Tco) and cold tolerance (ability to maintain Tco when exposed to 10 degrees C for 3 h) described for C57BL/6J mice are compared to other mouse strains. Assessment of young and aged male mice of the C57BL/6J and A/J inbred strains and their F1 hybrid, B6AF1/J, as well as a pen-bred strain of Mus musculus captured from the wild revealed an aged-related decline in thermoregulation among all these strains. The degree of decline in thermoregulation was roughly correlated to differences in strain-specific lifespan. Aged mice of the relatively short-lived genotype, A/J (mean lifespan of 22 months), had the lowest Tco and poorest cold tolerance. The long-lived hybrids, B6AF1/J (mean lifespan of 29 months), demonstrated the highest Tco and the best cold tolerance among aged mice. C57BL/6J (mean lifespan of 26 months) showed an intermediate level of thermoregulation. Aged pen-bred mice demonstrated a significant decline in cold tolerance, but not a significant decline in Tco. The thermoregulatory responses of the pen-bred mice were superior to those observed among the domesticated strains. These data suggest that the age-related impairment in thermoregulation is a general phenomenon of aging in Mus musculus that is correlated with strain-specific lifespan but is possibly affected by the level of hybridization and domestication.

Aging↗

ATP dependence of Na(+)-K+ pump of cold-sensitive and cold-tolerant mammalian red blood cells.

1. The ATP concentration of intact, cold-tolerant (ground squirrel) red cells and cold-sensitive (guinea-pig and human) red cells was monitored by use of the firefly tail, luciferin-luciferase assay. ATP kinetics of the pump in intact red blood cells was investigated by altering cell [ATP] by progressive depletion of ATP in the presence of 2-deoxy-D-glucose and then by measurement of ouabain-sensitive K+ influx at each level of [ATP] at various temperatures between 37 and 5 degrees C. Na(+)-K(+)-ATPase activity of broken membranes was also determined in parallel experiments using ouabain-sensitive release of 32P from [gamma-32P]ATP as a measure of activity. 2. Without depletion, there is no immediate decrease in [ATP] of intact cold-sensitive cells at low temperature (5 degrees C) at times when there are marked differences in the activities of the Na(+)-K+ pump of cold-tolerant and cold-sensitive cells. 3. At 37 degrees C Na(+)-K(+)-ATPase of all three species exhibited two components of ATP dependence at 37 degrees C, one with high velocity, low affinity, the other with low velocity, high affinity. Affinities of both components rose with cooling. 4. A similar, two component pattern was observed in intact guinea-pig and human red cells at 37 degrees C, except that the segment corresponding to the high affinity component had an apparent Km (Michaelis-Menten constant) 3- to 4-fold higher than that of the broken membrane preparation. 5. Cooling intact guinea-pig and human red cells decreased the apparent affinity of the high velocity, low affinity component for ATP, so that at 20 degrees C the value of Km approached or exceeded the levels of physiological ATP concentration. Below 20 degrees C only one component with values corresponding to that of the low velocity, high affinity component could be observed. 6. In intact ground squirrel cells only the low affinity, high velocity component was apparent between 37 and 5 degrees C. Its affinity for ATP rose with cooling between 37 and 5 degrees C.

Adenosine Triphosphate↗

Transformation of Skeletal Muscle from Fast- to Slow-Twitch during Acquisition of Cold Tolerance in the Chick.

Although birds lack brown adipose tissue, a thermogenic organ found in mammals, they possess other thermogenic mechanisms. In the current studies, we examined the molecular mechanisms of avian thermogenesis by studying how chicks acquire cold tolerance. We found that the acquisition of cold tolerance corresponded with an increase in the redness of the skeletal muscle, suggesting an increase in slow-twitch muscle fiber. This was confirmed by histological analysis. In addition, in chicks acquiring cold tolerance, there was an enhanced expression of the chicken homologue of peroxisome proliferator-activated receptor-gamma coactivator-1alpha, a protein involved in adaptive thermogenesis in mammalian brown adipose tissue and in slow-twitch fiber formation in mammalian skeletal muscle. Subtraction and differential display techniques further showed that, when chicks acquired cold tolerance, the expression of genes associated with slow-twitch fibers increased, whereas those associated with fast-twitch fibers decreased. There was also an enhanced expression of mitochondrial oxidative genes. Together, these results suggest that transformation of skeletal muscle fiber from fast-twitch to slow-twitch is involved in the acquisition of thermogenesis in chicks.

Adaptation, Physiological↗

Beyond the lipid hypothesis: mechanisms underlying phenotypic plasticity in inducible cold tolerance.

The physiological adjustment of organisms in response to temperature variation is a crucial part of coping with environmental stress. An important component of the cold response is the increase in membrane lipid unsaturation, and this has been linked to an enhanced resistance to the debilitating or lethal effects of cold. Underpinning the lipid response is the upregulation of fatty acid desaturases (des), particularly those introducing double bonds at the 9-10 position of saturated fatty acids. For plants and microbes there is good genetic evidence that regulation of des genes, and the consequent changes in lipid saturation, are causally linked to generation of a cold-tolerant phenotype. In animals, however, supporting evidence is almost entirely limited to correlations of saturation with cold conditions. We describe our recent attempts to provide a direct test of this relationship by genetic manipulation of the nematode Caenorhabditis elegans. We show that this species displays a strong cold tolerant phenotype induced by prior conditioning to cold, and that this is directly linked to upregulated des activity. However, whilst genetic disruption of des activity and lipid unsaturation significantly reduced cold tolerance, animals retained a substantial component of their stress tolerant phenotype produced by cold conditioning. This indicates that mechanisms other than lipid unsaturation play an important role in cold adaptation.

Acclimatization↗

Effects of starvation on body composition and cold tolerance in the collembolan Orchesella cincta and the isopod Porcellio scaber.

The effects of long-term starvation on the body composition of the isopod Porcellio scaber (Latreille) and the collembolan Orchesella cincta (L.) were studied, by determining the body composition in starved and fed animals. A period under summer conditions (19 degrees C, 75% RH and L/D 16/8 photoperiod), was followed by a period under winter conditions (5 degrees C, 75% RH and LD 6/18 photoperiod). O. cincta was held under summer conditions for 3weeks, during which its protein and lipid content decreased, while its water content increased. In P. scaber, the same occurred during the 6weeks they were kept under summer conditions. During subsequent weeks under winter conditions, changes in cold tolerance of the animals were investigated. Cold tolerance and haemolymph osmolality were measured once a week. Starved animals had lower cold tolerance than fed ones. For P. scaber a decreased haemolymph osmolality was found in starved animals compared to fed ones. This is assumed to be caused by a combination of the consumption of carbohydrates out of the haemolymph and of protein reserves and the accumulation of body water. O. cincta appeared to be capable of osmoregulation, as haemolymph osmolality did not differ between starved and fed animals, despite differences in body water content. Decreased cold tolerance in starved animals of both species may be caused by increased water content or, more probably, by the decrease in reserves needed to produce cryoprotective substances.

Journal Article↗

Snow white and the seven dwarfs: a multivariate approach to classification of cold tolerance.

Two main cold hardiness strategies of insects - freeze tolerance in some species, and overwintering in a supercooled state without tolerance of freezing in many others - were recently reclassified. However, I present several problems with the current systems. My suggested classification is based on clearer definitions of the causes of cold injury. I recognize three main mortality factors: freezing of body liquids, cold shock, and cumulative chill injury. Presence or absence of each of these factors produce eight combinations. I have named the eight classes after Snow White and the Seven Dwarfs to avoid nomenclatural confusion. Some of these classes are probably not used as tactics against cold injury by any insect species. Other classes contain so many species that they might be reclassified in more detail, using values of supercooling point and other quantitative parameters. However, widely comparable parameters, like the upper limit of cold injury zone and the sum of injurious temperatures are still rarely published, thus we still lack comprehensive data for multivariate analyses. Every cold hardiness strategy should be characterized by a meaningful class or subclass together with the physiological, biochemical, and behavioural mechanisms employed by the insects. I also point out the existence of strategies that combine two tactics - either a switching strategy (during preparation for winter, population "chooses" which tactic will be used), or a dual strategy (individuals are ready to use one of the tactics depending on the prevailing environmental conditions).

Journal Article↗

The effects of two types of clothing on seasonal cold tolerance.

This study investigated the effects of two types of clothing, leaving legs covered or uncovered, on seasonal cold tolerance in women. Experiments were carried out to compare cold tolerance at an ambient temperature (Ta) of 10 degrees C in December between two groups of subjects, who wore either knee-length skirts (skirt group) or full-length trousers (trouser group) for 3 months from September to November. The main results are summarized as follows: rectal temperatures continued to fall for 40 min in the trouser group when the subjects were covered by a blanket, while it became stable in 30 min in the skirt group; rectal temperatures showed greater increases in the skirt group when the blanket was removed after 40 min exposure to Ta of 10 degrees C; metabolic heat production was kept significantly lower in the skirt group when uncovered or covered by a blanket at Ta of 10 degrees C; metabolic heat production was negatively correlated with mean skin temperature and was always higher in the trouser group when measured at the same mean skin temperature; in the uncovered condition diastolic blood pressure increased significantly in the trouser group but not in the skirt group. These results would suggest that the subjects who wore skirts for 3 months from September to November had improved their ability to tolerate the cold.

Acclimatization↗

Effect of reproductive function on cold tolerance in deer mice.

Thermoregulatory responses were evaluated in male deer mice (Peromyscus maniculatus nebrascensis) after exposure to short photoperiod and either warm or cold ambient temperature (T(a)). Deer mice were chosen for this study because males exhibit differential reproductive responses to short day length (SD); this difference has a genetic basis, and both phenotypes are found within natural breeding populations. Deer mice undergoing SD-induced gonadal regression significantly improved their cold limit to -32.9 degrees C after exposure to SD/warm T(a) and to -47.4 degrees C after SD/cold T(a) exposure, relative to long day length/warm T(a) controls (-17.4 degrees C). In contrast, deer mice maintaining reproductive function despite SD exposure significantly improved cold limit to -27.2 degrees C only after exposure to SD/cold T(a), relative to controls (-16.3 degrees C). Maximum norepinephrine-induced nonshivering thermogenesis (NST) did not vary with reproductive state, indicating differences in cold tolerance were not due to capacity to produce heat by NST. Comparison between phenotypes of heat production during cold tolerance tests indicated that greater cold tolerance among mice exhibiting SD-induced gonadal regression can be accounted for by 1) lower rates of heat loss and 2) greater improvement of heat production. These findings suggest a functional relationship between reproductive function and seasonal thermoregulatory adjustments and indicate a significant cost to breeding during the winter months.

Adaptation, Physiological↗

Cold tolerance in hatchling painted turtles (Chrysemys picta): supercooling or tolerance for freezing?

We studied tolerance for cold in hatchling painted turtles (Chrysemys picta) from Lake Metigoshe, Bottineau County, North Dakota, to determine whether neonates in populations near the northern limit of distribution rely on a tolerance for freezing or on a capacity for supercooling to survive their first winter of life. We placed hatchlings individually into artificial hibernacula constructed in jars of damp, loamy sand and then cooled the jars to approximately -0.45 degrees C, which was below the equilibrium freezing point for water held by the sand but above that for body fluids of the neonatal turtles. A piece of ice next was placed on the surface of the sand in each jar to induce freezing of the soil water. After the soil water had frozen to an equilibrium, the temperature in the jars was lowered by 1 degrees C/d to minima averaging -2.5 degrees C, -4.5 degrees C, -6.5 degrees C, and -10.5 degrees C in different treatments. These temperatures were maintained for varying periods, so that animals in each treatment were exposed to temperatures below the equilibrium freezing point for their body fluids for a total of 11 d. Thirty of 32 hatchlings survived exposure to -2.5 degrees C; 24 of 32 survived at -4.5 degrees C; 14 of 32 withstood -6.5 degrees C; and 7 of 32 tolerated -10.5 degrees C. Freezing exotherms were detected in temperature profiles for turtles that succumbed but not in those for hatchlings that survived. Thus, the ability of hatchlings to withstand subzero temperatures for extended periods apparently requires that they avoid freezing. Although other workers contend that tolerance for freezing is the key to survival over winter by hatchling painted turtles from the region of Lake Metigoshe, our findings indicate that neonates rely primarily on their ability to remain unfrozen and supercooled.

Animals↗

Lasting effect of infantile cold experience on cold tolerance in adult rats.

The effect of short and repetitive exposure to cold (5 degrees C, 4 hr/day for 2 weeks) from the birth up to the 14th day of newborn rats onthe thermal regulation in adulthood and on the tolerance to cold was investigated. After being exposed to cold, they were transferred to a room at 25 degrees C (N-CA). The control rats were raised at 25 degrees C (N-WA). An acute cold exposure test was performed by placing the animals in a room at 5 degrees C under urethane anesthesia. Electrical activity of neck muscles as an index of shivering was recorded. The colonic temperature fell at a significantly slower rate in N-CA rats with less shivering than in N-WA ones. Nonshivering thermogenesis tested by norepinephrine was significantly greater in N-CA rats than in N-WA ones. These results suggest that N-CA rats developed improved cold tolerance accompanied by greater nonshivering thermogenesis. Such a phenomenon in N-CA lasted for 18 weeks after the termination of cold exposure. Adult rats subjected to the same scheme of cold exposure (A-CA) (5 degrees C, 4 hr/day, 2 weeks) showed essentially the same results as seen in N-CA, but its improved cold tolerance and elevated nonshivering thermogenesis disappeared 4 weeks after the termination of cold exposure. Extirpation of interscapular brown adipose tissue immediately before the cold test did not appreciably affect the cold tolerance in N-CA and A-CA rats. The colonic temperature at the onset of shivering was significantly lower in N-CA as well as A-CA rats than in each of the corresponding control rats, indicating a shift of the shivering threshold to lower temperature values in the animals exposed intermittently to cold. These results indicate that an infantile experience with cold results in a greater and longer sustained ability to tolerate cold in adulthood, characterized by enhanced nonshivering thermogenesis.

Acclimatization↗

QTLs conferring cold tolerance at the booting stage of rice using recombinant inbred lines from a japonica x indica cross.

Low temperature stress is common for rice grown in temperate regions and at high elevations in the tropics. The most senstive stage to this stress is booting, about 11 days before heading. Japonica cultivars are known to be more tolerant than indicas. We constructed a genetic map using 191 recombinant inbred lines derived from a cross between a temperate japonica, M-202, and a tropical indica, IR50, in order to locate quantitative trait loci (QTLs) conferring cold tolerance. The map with a total length of 1,276.8 cM and an average density of one marker every 7.1 cM was developed from 181 loci produced by 175 microsatellite markers. Cold tolerance was measured as the degree of spikelet sterility of treated plants at a 12 degrees C temperature for 5 days in the growth chamber. QTLs on chromosomes 1, 2, 3, 5, 6, 7, 9 and 12 were identified to confer cold tolerance at the booting stage. The QTL contribution to the phenotypic variation ranged from 11 to 17%. The two QTLs with the highest contribution to variation, designated qCTB2a and qCTB3, were derived from the tolerant parent, M-202, each explaining approximately 17% of the phenotypic variance. Two of the eight QTLs for cold tolerance were contributed by IR50.

Chromosome Mapping↗

[Evaluation of morphological traits on near-isogenic lines of cold tolerance and molecular validation at booting stage in Japonica rice].

Five parents and their near-isogenic lines (NILs) were grown in field experiments in Kunming (elevation 1916 m, two kinds of water temperatures at booting stage were 19.5 +/- 0.7 degrees C and 17.8 +/- 1.1 degrees C) and Aziying (elevation 2150 m, water temperature at booting stage was 18.2 +/- 0.22 degrees C), Yunnan, China, in 2004. Seventeen agronomic traits and SSR molecular markers were clustered by Statistica. The results showed that: (1) The morphological traits of NILs of cold tolerance and recurrent parents (Towada) at booting stage were similar, but the traits related cold tolerance, such as spike neck length, full grains, seed setting rate, anther length, anther volume, were different obviously; (2) using 7 markers had polymorphism between Towada and NILs were chosen from 78 SSR markers, the cold tolerance of NILs proved to be different to Towada. This demonstrates that these NILs were promising population for fine mapping and cloning genes for cold tolerance.

Breeding↗

Solubilization of inorganic phosphate and plant growth promotion by cold tolerant mutants of Pseudomonasfluorescens.

A study for screening and selection of cold tolerant mutants of Pseudomonas fluorescens strains GRS1, PRS9 and ATCC13525 based on 'P' solubilization ability and subsequent effect on plant growth promotion under in vitro and in situ conditions was conducted. Of all the mutants tested, two were selected, as there was a 21-fold increase in CRPF, (GRS, mutant) and a 10-fold decrease in CRPF7 (PRS9 mutant) over their respective wild types. Under in vitro conditions at 10 degrees C, these cold tolerant mutants exhibited increased plant growth indicating their functionality at low temperature. Subsequently, greenhouse trials using soil-plant microcosms were conducted which revealed that CRPF, (high 'P' solubilizer) was a good rhizosphere colonizer showing a significant increase in root (30 and 20%) and shoot length (20 and 24%) of mung bean, both in sterilized and unsterilized soil, respectively. On the contrary, CRPF, (low 'P' solubilizer) did not stimulate plant growth. Furthermore, sand experiments indicated that tricalcium phosphate served as better phosphorus source for CRPF2 treated mung bean seeds.

Cold Temperature↗

The effect of cold acclimation and deacclimation on cold tolerance, trehalose and free amino acid levels in Sitophilus granarius and Cryptolestes ferrugineus (Coleoptera).

Canadian and French laboratory strains of Sitophilus granarius (L.) and Cryptolestes ferrugineus (Stephens) were cold acclimated by placing adults at 15, 10 and 5 degrees C successively for 2wk at each temperature before deacclimating them for 1wk at 30 degrees C. Unacclimated S. granarius had an LT(50) (lethal time for 50% of the population) of 12days at 0 degrees C compared with 40days after the full cold acclimation. At -10 degrees C, unacclimated C. ferrugineus had an LT(50) of 1.4days compared with 24days after the full acclimation. Cold acclimation was lost within a week after returning insects to 30 degrees C. Trehalose, as well as the amino acids proline, asparagine, glutamic acid and lysine were higher in cold acclimated insects for both species. For S. granarius, glutamine was higher in cold acclimated insects and isoleucine, ethanolamine and phosphoethanolamine, a precursor of phospholipids, were lower in cold acclimated insects. For C. ferrugineus, alanine, aspartic acid, threonine, valine, isoleucine, leucine, phenylalanine and phosphoethanolamine were higher in cold acclimated insects. For both species tyrosine was lower in cold acclimated insects. There were small but significant differences between Canadian and French strains of S. granarius, with the Canadian strain being more cold hardy and having higher levels of trehalose. There were small but significant differences between male and female S. granarius, with males being more cold hardy and having higher levels of proline, asparagine and glutamic acid. In conclusion, high levels of trehalose and proline were correlated with cold tolerance, as seen in several other insects. However, correlation does not prove that these compounds are responsible for cold tolerance, and we outline further tests that could demonstrate a causal relationship between trehalose and proline and cold tolerance.

Journal Article↗

Neuronal deletion of Lepr elicits diabesity in mice without affecting cold tolerance or fertility.

Leptin signaling in the brain regulates energy intake and expenditure. To test the degree of functional neuronal leptin signaling required for the maintenance of body composition, fertility, and cold tolerance, transgenic mice expressing Cre in neurons (CaMKIIalpha-Cre) were crossed to mice carrying a floxed leptin receptor (Lepr) allele to generate mice with neuron-specific deletion of Lepr in approximately 50% (C F/F mice) and approximately 75% (C Delta17/F mice) of hypothalamic neurons. Leptin receptor (LEPR)-deficient mice (Delta17/Delta17) with heat-shock-Cre-mediated global Lepr deletion served as obese controls. At 16 wk, male C F/F, C Delta17/F, and Delta17/Delta17 mice were 13.2 (P < 0.05), 45.0, and 55.9% (P < 0.001) heavier, respectively, than lean controls, whereas females showed 31.6, 68.8, and 160.7% increases in body mass (P < 0.001). Significant increases in total fat mass (C F/F: P < 0.01; C Delta17/F and Delta17/Delta17:P < 0.001 vs. sex-matched, lean controls), and serum leptin concentrations (P < 0.001 vs. controls) were present in proportion to Lepr deletion. Male C Delta17/F mice had significant elevations in basal serum insulin concentrations (P < 0.001 vs. controls) and were glucose intolerant, as measured by glucose tolerance test (AUC P < 0.01 vs. controls). In contrast with previous observations in mice null for LEPR signaling, C F/F and C Delta17/F mice were fertile and cold tolerant. These findings support the hypothesis that body weight, adiposity, serum leptin concentrations, and glucose intolerance are proportional to hypothalamic LEPR deficiency. However, fertility and cold tolerance remain intact unless hypothalamic LEPR deficiency is complete.

Adaptation, Physiological↗

Effects of feeding beef females supplemental fat during gestation on cold tolerance in newborn calves.

Effects of prepartum fat supplementation of the dam on cold tolerance of calves were determined in two studies. In Exp. 1, 22 F1, crossbred heifers gestating F2 calves received diets containing either 1.7 or 4.7% dietary fat starting at d 230+/-2d of gestation. Safflower seeds (Carthamus tinctorius) containing 37% oil with 79% linoleic acid were the supplemental fat source in isocaloric-isonitrogenous diets. Calves were separated from their dams at birth, fed pooled dairy-cow colostrum, muzzled to prevent sucking, and returned to their dams in a heated (22 degrees C) barn for 3.5 h. At 4 h of age, a jugular catheter was inserted. At 5 h of age, calves were placed in a 0 degrees C room for 140 min and rectal temperatures and blood samples were obtained at 10- and 20-min intervals. Blood was assayed for glucose, cortisol, and cholesterol. In Exp. 2, 18 multiparous, crossbred beef cows bred to Murray Grey sires were randomly assigned to receive diets containing either 1.7 or 3.1% dietary fat starting at 235+/-2 d gestation. Safflower seeds were used as the supplemental fat source in isocaloric-isonitrogenous diets. At d 260 of gestation, premature parturition was induced in one-half of the cows from each diet group by feeding Ponderosa pine (Pinus ponderosa) needles. Experimental protocols were the same as in Exp. 1, except that cold exposure was at 9 degrees C for 200 min. Rectal temperatures were affected in Exp. 1 by time and diet x time (both P < .01) and diet x calf sex (P < .05) and in Exp. 2 by calf age (P < .05), time, and calf age x time (both P < .01). Plasma cortisol concentrations were affected by time (P < .01) and calf sex x time (P < .05) in Exp. 1 and by time ( P < .01) in Exp. 2. Cholesterol concentrations in Exp. 1 were affected by diet x time (P < .05) and in Exp. 2 by time (P < .05). Plasma glucose concentrations were affected in Exp. 1 by diet (P < .05) and in Exp. 2 by calf age, time, and calf age x time (all P < .01). We conclude from Exp. 1 that feeding heifers supplemental fat during late gestation increased glucose concentrations in the newborn calf, resulting in favorable responses in body temperature in the cold-stressed newborns. This increase in substrate availability suggests a potential positive effect on heat generation in newborns during sustained periods of cold stress. In Exp. 2, premature calves had compromised cold tolerance possibly due to impaired shivering or brown adipose tissue thermogenesis.

Adaptation, Physiological↗

Cold-acclimation improves cold-tolerance of diabetic rats.

1. The aim of these experiments was to study the extent to which previous cold-acclimation improves the cold-tolerance of diabetic rats. 2. Alloxan diabetic rats (fasting blood glucose higher than 200 mg/dl) were used in the experiments. 3. In Expt. 1, non-cold-acclimated control and diabetic rats were exposed to cold environment (7-9 degrees C), and the percentage of survival calculated during a 12-day experimental period. In Expt. 2, the rats were previously cold-acclimated before alloxan or saline injection (diabetic and control cold-acclimated rats) and the survival rate was also assessed during a 12-day period in the cold. 4. The percentage of survival of the non-cold-acclimated diabetic rats (Expt.1) was 19% compared with 79% of the diabetic cold-acclimated animals (Expt. 2). There were no deaths in the control groups. 5. Cold-acclimated diabetic rats maintained a near-normal thermogenic response after noradrenaline injection. This response was impaired in non-cold-acclimated diabetic rats. 6. The results of these experiments suggest that the enhanced cold-tolerance of diabetic cold-acclimated rats could be related to the increased sympathetic activity and enhanced insulin sensitivity in thermogenic tissues, such as brown fat.

Acclimatization↗