Paula Redman

Uncoupled and surviving: individual mice with high metabolism have greater mitochondrial uncoupling and live longer

Two theories of how energy metabolism should be associated with longevity, both mediated via free‐radical production, make completely contrary predictions. The ‘rate of living‐free‐radical theory’ ( Pearl, 1928 ; Harman, 1956 ; Sohal, 2002 ) suggests a negative association, the ‘uncoupling to survive’ hypothesis ( Brand, 2000 ) suggests the correlation should be positive. Existing empirical data on this issue is contradictory and extremely confused ( Rubner, 1908 ; Yan & Sohal, 2000 ; Ragland & Sohal, 1975 ; Daan et al., 1996 ; Wolf & Schmid‐Hempel, 1989 ]. We sought associations between longevity and individual variations in energy metabolism in a cohort of outbred mice. We found a positive association between metabolic intensity (kJ daily food assimilation expressed as g/body mass) and lifespan, but no relationships of lifespan to body mass, fat mass or lean body mass. Mice in the upper quartile of metabolic intensities had greater resting oxygen consumption by 17% and lived 36% longer than mice in the lowest intensity quartile. Mitochondria isolated from the skeletal muscle of mice in the upper quartile had higher proton conductance than mitochondria from mice from the lowest quartile. The higher conductance was caused by higher levels of endogenous activators of proton leak through the adenine nucleotide translocase and uncoupling protein‐3. Individuals with high metabolism were therefore more uncoupled, had greater resting and total daily energy expenditures and survived longest – supporting the ‘uncoupling to survive’ hypothesis.

Life-long vitamin C supplementation in combination with cold exposure does not affect oxidative damage or lifespan in mice, but decreases expression of antioxidant protection genes

Oxidative stress is suggested to be central to the ageing process, with endogenous antioxidant defence and repair mechanisms in place to minimize damage. Theoretically, supplementation with exogenous antioxidants might support the endogenous antioxidant system, thereby reducing oxidative damage, ageing-related functional decline and prolonging life- and health-span. Yet supplementation trials with antioxidants in animal models have had minimal success. Human epidemiological data are similarly unimpressive, leading some to question whether vitamin C, for example, might have pro-oxidant properties in vivo. We supplemented cold exposed (7+/-2 degrees C) female C57BL/6 mice over their lifespan with vitamin C (ascorbyl-2-polyphosphate), widely advocated and self administered to reduce oxidative stress, retard ageing and increase healthy lifespan. No effect on mean or maximum lifespan following vitamin C treatment or any significant impact on body mass, or on parameters of energy metabolism was observed. Moreover, no differences in hepatocyte and lymphocyte DNA oxidative damage or hepatic lipid peroxidation was seen between supplemented and control mice. Using a DNA macroarray specific for oxidative stress-related genes, we found that after 18 months of supplementation, mice exhibited a significantly reduced expression of several genes in the liver linked to free-radical scavenging, including Mn-superoxide dismutase. We confirmed these effects by Northern blotting and found additional down-regulation of glutathione peroxidase (not present on macroarray) in the vitamin C treated group. We suggest that high dietary doses of vitamin C are ineffective at prolonging lifespan in mice because any positive benefits derived as an antioxidant are offset by compensatory reductions in endogenous protection mechanisms, leading to no net reduction in accumulated oxidative damage.

Effect of photoperiod on body mass, food intake and body composition in the field vole, Microtus agrestis.

SUMMARY Many small mammals respond to seasonal changes in photoperiod by altering body mass and adiposity. These animals may provide valuable models for understanding the regulation of energy balance. Here, we present data on the field vole (Microtus agrestis) – a previously uncharacterised example of photoperiod-induced changes in body mass. We examined the effect of increased day length on body mass, food intake, apparent digestive efficiency, body composition, de novo lipogenesis and fatty acid composition of adipose tissue in cold-acclimated (8°C) male field voles by transferring them from a short (SD, 8 h:16 h L:D) to long day photoperiod (LD, 16 h:8 h L:D). During the first 4 weeks of exposure to LD, voles underwent a substantial increase in body mass, after which the average difference between body masses of LD and SD voles stabilized at 7.5 g. This 24.8% increase in body mass reflected significant increases in absolute amounts of all body components, including dry fat mass, dry lean mass and body water mass. After correcting body composition and organ morphology data for the differences in body mass, only gonads (testes and seminal vesicles) were enlarged due to photoperiod treatment. To meet energetic demands of deposition and maintenance of extra tissue, voles adjusted their food intake to an increasing body mass and improved their apparent digestive efficiency. Consequently, although mass-corrected food intake did not differ between the photoperiod groups, the LD voles undergoing body mass increase assimilated on average 8.4 kJ day-1 more than animals maintained in SD. The majority (73–77%) of the fat accumulated as adipose tissue had dietary origin. The rate of de novo lipogenesis and fatty acid composition of adipose tissue were not affected by photoperiod. The most important characteristics of the photoperiodic regulation of energy balance in the field vole are the clear delineation between phases where animals regulate body mass at two different levels and the rate at which animals are able to switch between different levels of energy homeostasis. Our data indicate that the field vole may provide an attractive novel animal model for investigation of the regulation of body mass and energy homeostasis at both organism and molecular levels.

Trade-offs between activity and thermoregulation in a small carnivore, the least weasel Mustela nivalis

We studied factors influencing daily energy expenditures (DEE) of male least weasels (Mustela nivalis) using the doubly labelled water technique. The relationship between ambient temperature and DEE formed a triangular pattern, characterized by invariance of the maximum DEE and an inverse relationship between minimum DEE and temperature. A simple energetic model relating the DEE of male weasels to activity time (AT) and ambient temperature predicted that, across seasons, less than 10 per cent of measurements approach the upper bound of observed DEE. Male weasels were able to maintain a relatively constant maximum energy output across varying temperatures by adjusting their AT to changes in temperature. They achieved maximum energy expenditures in winter due to high thermoregulatory costs, and in spring and summer due to high levels of physical activity. This pattern exemplifies a ‘metabolic niche’ of a small mammal having extremely high energy expenditures primarily driven by ambient temperature.

Lifelongα-tocopherol supplementation increases the median life span of C57BL/6 mice in the cold but has only minor effects on oxidative damage

The effects of dietary antioxidant supplementation on oxidative stress and life span are confused. We maintained C57BL/6 mice at 7 +/- 2 degrees C and supplemented their diet with alpha-tocopherol from 4 months of age. Supplementation significantly increased (p = 0.042) median life span by 15% (785 days, n = 44) relative to unsupplemented controls (682 days, n = 43) and also increased maximum life span (oldest 10%, p = 0.028). No sex or sex by treatment interaction effects were observed on life span, with treatment having no effect on resting or daily metabolic rate. Lymphocyte and hepatocyte oxidative DNA damage and hepatic lipid peroxidation were unaffected by supplementation, but hepatic oxidative DNA damage increased with age. Using a cDNA macroarray, genes associated with xenobiotic metabolism were significantly upregulated in the livers of female mice at 6 months of age (2 months supplementation). At 22 months of age (18 months supplementation) this response had largely abated, but various genes linked to the p21 signaling pathway were upregulated at this time. We suggest that alpha-tocopherol may initially be metabolized as a xenobiotic, potentially explaining why previous studies observe a life span extension generally when lifelong supplementation is initiated early in life. The absence of any significant effect on oxidative damage suggests that the life span extension observed was not mediated via any antioxidant properties of alpha-tocopherol. We propose that the life span extension observed following alpha-tocopherol supplementation may be mediated via upregulation of cytochrome p450 genes after 2 months of supplementation and/or upregulation of p21 signaling genes after 18 months of supplementation. However, these signaling pathways now require further investigation to establish their exact role in life span extension following alpha-tocopherol supplementation.

Male short-tailed field voles (Microtus agrestis) build better insulated nests than females.

Abstract Nest construction is an extremely widespread behaviour. In small endotherms the nest serves primarily to provide insulation, and thereby retard heat loss of the constructor, or its offspring. In arctic and temperate regions many small mammals build nests to protect themselves from low ambient temperatures. We measured the physical properties of nests built by short-tailed field voles Microtus agrestis that were kept in captivity under cold conditions. The most important factor influencing nest insulation was nest wall thickness; however, nests with thick walls also contained more nesting material. Insulative capacity of the nest did not reach an asymptote up to nests containing 20 g of material. Nest insulation was not correlated with resting metabolic rate, body mass or body composition of the vole that constructed the nest. However, nests built by males had greater insulation than those made by females; males also had significantly lower food intake rates when compared to females with nests. No significant difference was observed in either fat mass or whole animal thermal conductance between males and females. Thermal conductance did increase significantly with increasing body mass, although not with resting metabolic rate. Voles with nests for prolonged periods had lower food intakes than voles without nests. The absolute saving averaged 1.9 g and was independent of body mass. This was a 28% saving on intake for a 22-g vole but only an 18% saving for a 40-g individual. When voles had nests for short periods they used the energy they saved to reduce food intake and increase body mass.

Determining Seabird Body Condition Using Nonlethal Measures

Energy stores are critical for successful breeding, and longitudinal studies require nonlethal methods to measure energy stores (“body condition”). Nonlethal techniques for measuring energy reserves are seldom verified independently. We compare body mass, size-corrected mass (SCM), plasma lipids, and isotopic dilution with extracted total body lipid content in three seabird species (thick-billed murres Uria lomvia, all four measures; northern fulmars Fulmarus glacialis, three measures; and black-legged kittiwakes Rissa tridactyla, two measures). SCM and body mass were better predictors of total body lipids for the species with high percent lipids (fulmars; ) than for the species with low percent lipids (murres and kittiwakes; ). The relationship between SCM and percent body lipids, which we argue is often a better measure of condition, was also poor () for species with low lipids. In a literature comparison of 17 bird species, percent lipids was the only predictor of the strength of the relationship between mass and total body lipids; we suggest that SCM be used as an index of energy stores only when lipids exceed 15% of body mass. Across all three species we measured, SCM based on the ordinary least squares regression of mass on the first principal component outperformed other measures. Isotopic dilution was a better predictor of both total body lipids and percent body lipids than were mass, SCM, or plasma lipids in murres. Total body lipids decreased through the breeding season at both sites, while total and neutral plasma lipid concentrations increased at one site but not another, suggesting mobilization of lipid stores for breeding. A literature review showed substantial variation in the reliability of plasma markers, and we recommend isotopic dilution (oxygen-18, plateau) for determination of energy reserves in birds where lipid content is below 15%.

Energy metabolism of Inuit sled dogs.

We explored how seasonal changes in temperature, exercise and food supply affected energy metabolism and heart rate of Inuit dogs in Greenland. Using open flow respirometry, doubly labeled water, and heart rate recording, we measured metabolic rates of the same dogs at two different locations: at one location the dogs were fed with high energy food throughout the year while at the other location they were fed with low energy food during summer. Our key questions were: is resting metabolic rate (RMR) increased during the winter season when dogs are working? Does feeding regime affect RMR during summer? What is the proportion of metabolic rate (MR) devoted to specific dynamic action (SDA), and what is the metabolic scope of working Inuit sled dogs? The Inuit dogs had an extremely wide thermoneutral zone extending down to −25°C. Temperature changes between summer and winter did not affect RMR, thus summer fasting periods were defined as baseline RMR. Relative to this baseline, summer MR was upregulated in the group of dogs receiving low energy food, whereas heart rate was downregulated. However, during food digestion, both MR and HR were twice their respective baseline values. A continuously elevated MR was observed during winter. Because temperature effects were excluded and because there were also no effects of training, we attribute winter elevated MR to SDA because of the continuous food supply. Working MR during winter was 7.9 times the MR of resting dogs in winter, or 12.2 times baseline MR.

Energy expenditure and water turnover in hunting dogs in winter conditions

Hunting with dogs in winter conditions is practised in the Nordic countries. The present study aimed at determining daily energy expenditure (DEE) and body water turnover (BWT) by the doubly labelled water technique in eight hunting dogs (body-weight (BW) range 14-27 kg) working 3 h/d for 3 d (-6 °C) on ground covered with 20-40 cm of loose snow, to provide information on energy and water requirements. The mean distance run during the hunting period was recorded by the global positioning system and averaged 19.4 km/d. DEE increased with increasing BW (P < 0.001) and varied between 7.20 and 16.6 MJ/d (mean 11.0 MJ/d) corresponding to 950-1350 kJ/kg BW(0.75) per d (mean 1170 kJ/kg BW(0.75) per d). The larger dogs tended to run longer than the smaller dogs and therefore spent more energy per kg BW(0.75) but not significantly (P>0.05). DEE values determined were close to the values measured for hunting dogs running for 3 h/d in hot climates, suggesting that climate within the range of the two studies has little impact on energy expenditure per h running activity. Compared with the work of sled dogs per km travelled running on a track, the work performed by the hunting dogs was suggested to be higher when running in a loose snow layer. However, DEE was much lower because sled dogs ran for a longer distance each day. Mean BWT was 217 ml/kg BW(0.75) or 19 ml/kJ metabolisable energy.

Effects of the doubly labelled water procedure on Great Tits Parus major feeding young

Capsule Female feeding rate behaviour was highly variable between individuals but no effect of the doubly labelled water (DLW) procedure on female visit rate was detected, whereas visit rates by males, which were neither trapped or manipulated, increased following trapping and manipulation of their partners. Aims To test the assumption that the subjects behaviour is normal during the measurement period when using DLW to measure energy expenditure. Methods Visits to the nestbox by parents feeding young were counted separately for females and males on the day before and the day after the female only was trapped to measure her energy expenditure using DLW. Visit rates were also counted for control pairs. Results Female visit rates did not differ before and after manipulation, or between experimental and control pairs, but bird behaviour was highly variable between individuals. In contrast, the visit rates of both experimental and control males, which were not trapped, increased on the second day. Conclusion The results for female behaviour supported the assumption of normality, but a small subset of particular individuals may be prone to adverse reactions. The response of the males may have been a reaction to disturbance at the nest.