Bronwyn M. McAllan

The degree of dietary fatty acid unsaturation affects torpor patterns and lipid composition of a hibernator

Diets rich in unsaturated and polyunsaturated fatty acids have a positive effect on mammalian torpor, whereas diets rich in saturated fatty acids have a negative effect. To determine whether the number of double bonds in dietary fatty acids are responsible for these alterations in torpor patterns, we investigated the effect of adding to the normal diet 5% pure fatty acids of identical chain length (C18) but a different number of double bonds (0, 1, or 2) on the pattern of hibernation of the yellow-pine chipmunk, Eutamias amoenus. The response of torpor bouts to a lowering of air temperature and the mean duration of torpor bouts at an air temperature of 0.5°C (stearic acid C18:0, 4.5±0.8 days, oleic acid C18:1, 8.6±0.5 days; linoleic acid C18:2, 8.5±0.7 days) differed among animals that were maintained on the three experimental diets. The mean minimum body temperatures (C18:0, +2.3±0.3°C; C18:1, +0.3±0.2°C; C18:2,-0.2±0.2°C), which torpid individuals defended by an increase in metabolic rate, and the metabolic rate of torpid animals also differed among diet groups. Moreover, diet-induced differences were observed in the composition of total lipid fatty acids from depot fat and the phospholipid fatty acids of cardiac mitochondria. For depot fat 7 of 13 and for heart mitochondria 7 of 14 of the identified fatty acids differed significantly among the three diet groups. Significant differences among diet groups were also observed for the sum of saturated, unsaturated and polyunsaturated fatty acids. These diet-induced alterations of body fatty acids were correlated with some of the diet-induced differences in variables of torpor. The results suggest that the degree of unsaturation of dietary fatty acids influences the composition of tissues and membranes which in turn may influence torpor patterns and thus survival of hibernation.

Coping with chaos: unpredictable food supplies intensify torpor use in an arid-zone marsupial, the fat-tailed dunnart (Sminthopsis crassicaudata).

The severity, duration and amplitude of extreme weather events are forecast to intensify with current climate trends, over both long (e.g. seasonal) and short (e.g. daily) time-scales. As such, the predictability of food supplies for many small endotherms is likely to become increasingly important. Numerous small mammals and birds combat food shortages using torpor, a controlled reduction in metabolic rate and body temperature that helps lower their daily energy requirements. As such, torpor often has been cited as a key feature allowing some small endotherms to survive highly unpredictable climates, such as tropics or dry deserts, but mensurative demonstrations of this are lacking. We have shown here that when a small desert marsupial, the fat-tailed dunnart (Sminthopsis crassicaudata), is offered unpredictable levels of daily food, they increase frequency of daily torpor and length of bouts compared with animals offered ad libitum food, but this was not found for animals offered a 70% food-restricted diet. Our data suggest that simple food restriction may not be sufficient for evaluating the efficacy of torpor as a strategy for managing unpredictable climates.

Torpor during Reproduction in Mammals and Birds: Dealing with an Energetic Conundrum

Torpor and reproduction in mammals and birds are widely viewed as mutually exclusive processes because of opposing energetic and hormonal demands. However, the reported number of heterothermic species that express torpor during reproduction is ever increasing, to some extent because of recent work on free-ranging animals. We summarize current knowledge about those heterothermic mammals that do not express torpor during reproduction and, in contrast, examine those heterothermic birds and mammals that do use torpor during reproduction. Incompatibility between torpor and reproduction occurs mainly in high-latitude sciurid and cricetid rodents, which live in strongly seasonal, but predictably productive habitats in summer. In contrast, torpor during incubation, brooding, pregnancy, or lactation occurs in nightjars, hummingbirds, echidnas, several marsupials, tenrecs, hedgehogs, bats, carnivores, mouse lemurs, and dormice. Animals that enter torpor during reproduction often are found in unpredictable habitats, in which seasonal availability of food can be cut short by changes in weather, or are species that reproduce fully or partially during winter. Moreover, animals that use torpor during the reproductive period have relatively low reproductive costs, are largely insectivorous, carnivorous, or nectarivorous, and thus rely on food that can be unpredictable or strongly seasonal. These species with relatively unpredictable food supplies must gain an advantage by using torpor during reproduction because the main cost is an extension of the reproductive period; the benefit is increased survival of parent and offspring, and thus fitness.

Expression of thyroid hormone transporters and deiodinases at the brain barriers in the embryonic chicken: Insights into the regulation of thyroid hormone availability during neurodevelopment.

Thyroid hormones (THs) are key regulators in the development of the vertebrate brain. Therefore, TH access to the developing brain needs to be strictly regulated. The brain barriers separate the central nervous system from the rest of the body and impose specific transport mechanisms on the exchange of molecules between the general circulation and the nervous system. As such they form ideal structures for regulating TH exchange between the blood and the brain. To investigate the mechanism by which the developing brain regulates TH availability, we investigated the ontogenetic expression profiles of TH transporters, deiodinases and the TH distributor protein transthyretin (TTR) at the brain barriers during embryonic and early postnatal development using the chicken as a model. In situ hybridisation revealed expression of the TH transporters monocarboxylate transporter 8 (MCT8) and 10 (MCT10), organic anion transporting polypeptide 1C1 (OATP1C1) and L-type amino acid transporter 1 (LAT1) and the inactivating type 3 deiodinase (D3) in the choroid plexus which forms the blood-cerebrospinal fluid barrier. This was confirmed by quantitative PCR which additionally indicated strongly increasing expression of TTR as well as detectable expression of the activating type 2 deiodinase (D2) and the (in)activating type 1 deiodinase (D1). In the brain capillaries forming the blood-brain barrier in situ hybridisation showed exclusive expression of LAT1 and D2. The combined presence of LAT1 and D2 in brain capillaries suggests that the blood-brain barrier forms the main route for receptor-active T3 uptake into the embryonic chicken brain. Expression of multiple transporters, deiodinases and TTR in the choroid plexus indicates that the blood-cerebrospinal fluid barrier is also important in regulating early TH availability. The impact of these barrier systems can be deduced from the clear difference in T3 and T4 levels as well as the T3/T4 ratio between the developing brain and the general circulation. We conclude that the tight regulation of TH exchange at the brain barriers from early embryonic stages is one of the factors needed to allow the brain to develop within a relative microenvironment.

The effects of cortisol and testosterone on renal function in male Antechinus stuartii (Marsupialia)

Abstract Seasonal changes in the physiology of Antechinus stuartii result in complete male mortality after mating. The most important endocrine changes in males are large rises in plasma testosterone and cortisol concentrations. Glomerular filtration rate (GFR) in males declines coincident with high plasma testosterone and cortisol. In the present study GFRs were measured in males captured in May (when endogenous plasma testosterone and cortisol levels are low) and given depot injections of either saline, testosterone-only, cortisol-only or testosterone plus cortisol at doses designed to mimic plasma levels during the mating period. GFR decreased significantly with testosterone injection, independent of cortisol treatment. Urinary concentrations of sodium and chloride, and osmolality decreased significantly with cortisol treatment, although the addition of testosterone reversed the effect. Total urinary excretion of electrolytes was similar between groups. Plasma potassium levels significantly increased in testosterone plus cortisol treated males. Plasma sodium levels significantly increased and plasma chloride significantly decreased in all groups treated with cortisol. Water consumption significantly increased in all cortisol-treated males and food consumption significantly increased in all testosterone-treated males. The seasonal renal functional changes observed in A. stuartii were mimicked by testosterone administration.

Phylogenetic differences of mammalian basal metabolic rate are not explained by mitochondrial basal proton leak.

Metabolic rates of mammals presumably increased during the evolution of endothermy, but molecular and cellular mechanisms underlying basal metabolic rate (BMR) are still not understood. It has been established that mitochondrial basal proton leak contributes significantly to BMR. Comparative studies among a diversity of eutherian mammals showed that BMR correlates with body mass and proton leak. Here, we studied BMR and mitochondrial basal proton leak in liver of various marsupial species. Surprisingly, we found that the mitochondrial proton leak was greater in marsupials than in eutherians, although marsupials have lower BMRs. To verify our finding, we kept similar-sized individuals of a marsupial opossum (Monodelphis domestica) and a eutherian rodent (Mesocricetus auratus) species under identical conditions, and directly compared BMR and basal proton leak. We confirmed an approximately 40 per cent lower mass specific BMR in the opossum although its proton leak was significantly higher (approx. 60%). We demonstrate that the increase in BMR during eutherian evolution is not based on a general increase in the mitochondrial proton leak, although there is a similar allometric relationship of proton leak and BMR within mammalian groups. The difference in proton leak between endothermic groups may assist in elucidating distinct metabolic and habitat requirements that have evolved during mammalian divergence.

Extensive production of Neospora caninum tissue cysts in a carnivorous marsupial succumbing to experimental neosporosis

Experimental infections of Sminthopsis crassicaudata, the fat-tailed dunnart, a carnivorous marsupial widely distributed throughout the arid and semi-arid zones of Australia, show that this species can act as an intermediate host for Neospora caninum. In contrast to existing models that develop relatively few N. caninum tissue cysts, dunnarts offer a new animal model in which active neosporosis is dominated by tissue cyst production. The results provide evidence for a sylvatic life cycle of N. caninum in Australia between marsupials and wild dogs. It establishes the foundation for an investigation of the impact and costs of neosporosis to wildlife.

A Functional Nexus between Photoperiod Acclimation, Torpor Expression and Somatic Fatty Acid Composition in a Heterothermic Mammal

The seasonal changes in thermal physiology and torpor expression of many heterothermic mammals are controlled by photoperiod. As function at low body temperatures during torpor requires changes of tissue lipid composition, we tested for the first time whether and how fatty acids are affected by photoperiod acclimation in hamsters, Phodopus sungorus, a strongly photoperiodic species. We also examined changes in fatty acid composition in relation to those in morphology and thermal biology. Hamsters in short photoperiod had smaller reproductive organs and most had a reduced body mass in comparison to those in long photoperiod. Pelage colour of hamsters under short photoperiod was almost white while that of long photoperiod hamsters was grey-brown and black. Short photoperiod acclimation resulted in regular (28% of days) torpor use, whereas all hamsters in long photoperiod remained normothermic. The composition of total fatty acids differed between acclimation groups for brown adipose tissue (5 of 8 fatty acids), heart muscle (4 of 7 fatty acids) and leg muscle (3 of 11 fatty acids). Importantly, 54% of all fatty acids detected were correlated (r2 = 0.60 to 0.87) with the minimum surface temperature of individuals, but the responses of tissues differed. While some of the compositional changes of fatty acids were consistent with a ‘homeoviscous’ response, this was not the case for all, including the sums of saturated and unsaturated fatty acids, which did not differ between acclimation groups. Our data identify a possible nexus between photoperiod acclimation, morphology, reproductive biology, thermal biology and fatty acid composition. They suggest that some of the changes in thermal physiology are linked to the composition of tissue and organ fatty acids.

Adaptation of phenylalanine and tyrosine catabolic pathway to hibernation in bats.

Some mammals hibernate in response to harsh environments. Although hibernating mammals may metabolize proteins, the nitrogen metabolic pathways commonly activated during hibernation are not fully characterized. In contrast to the hypothesis of amino acid preservation, we found evidence of amino acid metabolism as three of five key enzymes, including phenylalanine hydroxylase (PAH), homogentisate 1,2-dioxygenase (HGD), fumarylacetoacetase (FAH), involved in phenylalanine and tyrosine catabolism were co-upregulated during hibernation in two distantly related species of bats, Myotis ricketti and Rhinolophus ferrumequinum. In addition, the levels of phenylalanine in the livers of these bats were significantly decreased during hibernation. Because phenylalanine and tyrosine are both glucogenic and ketogenic, these results indicate the role of this catabolic pathway in energy supply. Since any deficiency in the catabolism of these two amino acids can cause accumulations of toxic metabolites, these results also suggest the detoxification role of these enzymes during hibernation. A higher selective constraint on PAH, HPD, and HGD in hibernators than in non-hibernators was observed, and hibernators had more conserved amino acid residues in each of these enzymes than non-hibernators. These conserved amino acid residues are mostly located in positions critical for the structure and activity of the enzymes. Taken together, results of this work provide novel insights in nitrogen metabolism and removal of harmful metabolites during bat hibernation.

The influence of reproductive hormones on the torpor patterns of the marsupial Sminthopsis macroura: Bet-hedging in an unpredictable environment

Seasonal cycles of reproduction are common in many mammals and these are combined with the necessary energy budgeting for thermoregulatory challenges. Many mammals meet the challenge of changing environmental temperatures in winter by using torpor, a controlled reduction in body temperature and metabolic rate. We aimed to determine the effects of photoperiod and reproductive hormones on the seasonal cycles of reproduction and torpor use in a marsupial that commences reproduction in winter, the stripe-faced dunnart, Sminthopsis macroura. Males and females were placed under LD 14:10 and natural reproductive hormones blocked by either flutamide (males) or mifepristone (females) or tamoxifen (females). Reproductive parameters, metabolic rate and torpor variables were determined. The same animals were then placed under LD 10:14 and given testosterone (males) or progesterone (females) or oestrogen (females). Reproductive parameters, metabolic rate and torpor variables were measured. Body mass and tail widths (fattening indicator) in males were significantly affected by testosterone, and the effects were reversed by hormone blockers. Reproductive parameters were unaffected. Resting metabolic rate and ability to use torpor were not affected by treatment in males, however torpor characteristics, especially torpor bout duration, were affected by presence of testosterone in males. In females, body mass was unaffected by hormone presence, although tail widths were affected. Disruption of reproductive cycles occurred with hormone blockers in females, however, resting metabolic rate was not affected, and only presence of progesterone affected torpor characteristics in females. Our results differ from those found for rodents, where presence of testosterone abolishes the use of torpor in males, and oestrogen inhibits torpor use in females. Our study suggests that, in this mammal, metabolic responses to the presence or absence of reproductive hormones differs between males and females, and there is no absolute endocrinologically-driven reproductive season demarcated from the torpor season.