Akt signalling through GSK-3beta, mTOR and Foxo1 is involved in human skeletal muscle hypertrophy and atrophy

Skeletal muscle size is tightly regulated by the synergy between anabolic and catabolic signalling pathways which, in humans, have not been well characterized. Akt has been suggested to play a pivotal role in the regulation of skeletal muscle hypertrophy and atrophy in rodents and cells. Here we measured the amount of phospho-Akt and several of its downstream anabolic targets (glycogen synthase kinase-3β (GSK-3β), mTOR, p70s6k and 4E-BP1) and catabolic targets (Foxo1, Foxo3, atrogin-1 and MuRF1). All measurements were performed in human quadriceps muscle biopsies taken after 8 weeks of both hypertrophy-stimulating resistance training and atrophy-stimulating de-training. Following resistance training a muscle hypertrophy (∼10%) and an increase in phospho-Akt, phospho-GSK-3β and phospho-mTOR protein content were observed. This was paralleled by a decrease in Foxo1 nuclear protein content. Following the de-training period a muscle atrophy (5%), relative to the post-training muscle size, a decrease in phospho-Akt and GSK-3β and an increase in Foxo1 were observed. Atrogin-1 and MuRF1 increased after the hypertrophy and decreased after the atrophy phases. We demonstrate, for the first time in human skeletal muscle, that the regulation of Akt and its downstream signalling pathways GSK-3β, mTOR and Foxo1 are associated with both the skeletal muscle hypertrophy and atrophy processes.

Water deprivation induces appetite and alters metabolic strategy in Notomys alexis : unique mechanisms for water production in the desert

Like many desert animals, the spinifex hopping mouse, Notomys alexis, can maintain water balance without drinking water. The role of the kidney in producing a small volume of highly concentrated urine has been well-documented, but little is known about the physiological mechanisms underpinning the metabolic production of water to offset obligatory water loss. In Notomys, we found that water deprivation (WD) induced a sustained high food intake that exceeded the pre-deprivation level, which was driven by parallel changes in plasma leptin and ghrelin and the expression of orexigenic and anorectic neuropeptide genes in the hypothalamus; these changed in a direction that would stimulate appetite. As the period of WD was prolonged, body fat disappeared but body mass increased gradually, which was attributed to hepatic glycogen storage. Switching metabolic strategy from lipids to carbohydrates would enhance metabolic water production per oxygen molecule, thus providing a mechanism to minimize respiratory water loss. The changes observed in appetite control and metabolic strategy in Notomys were absent or less prominent in laboratory mice. This study reveals novel mechanisms for appetite regulation and energy metabolism that could be essential for desert rodents to survive in xeric environments.

Foraging behaviour of two rodent species inhabiting a kopje (rocky outcrop) in Tsavo West National Park, Kenya

We employed patch use theory to evaluate how several environmental factors influence the foraging behaviour of two rodent species: Grammomys dolichurus and Acomys cahirinus. Foraging efficiency was determined by measuring the remaining food in artificial food patches (giving-up densities: GUDs) from two experiments. In the first experiment, we placed patches in different microhabitat types (cover vs open) and at varying distances from cover. This experiment was conducted during three moon stages (waxing, full, waning). We found that the rodents had higher GUDs (lower foraging efficiency) in the open microhabitat. The distance from nearest shelter had a marginally significant positive effect on GUDs. GUDs were higher in both microhabitat types during the waxing and full phases, but decreased sharply once the moon began to rise after sunset. These results are likely due to higher predation risk away from cover and in more illuminated environments. In the second experiment, we examined mouse responses to seeds impregnated with plant toxins. Seeds impregnated with oxalic acid were avoided by the rodents, while seeds soaked in tannic acid did not differ significantly from control seeds. Our results highlight important ecological factors affecting the foraging behaviour of these rodents.

Tyk2 and Stat3 regulate brown adipose tissue differentiation and obesity

Mice lacking the Jak tyrosine kinase member Tyk2 become progressively obese due to aberrant development of Myf5+ brown adipose tissue (BAT). Tyk2 RNA levels in BAT and skeletal muscle, which shares a common progenitor with BAT, are dramatically decreased in mice placed on a high-fat diet and in obese humans. Expression of Tyk2 or the constitutively active form of the transcription factor Stat3 (CAStat3) restores differentiation in Tyk2−/− brown preadipocytes. Furthermore, Tyk2−/− mice expressing CAStat3 transgene in BAT also show improved BAT development, normal levels of insulin, and significantly lower body weights. Stat3 binds to PRDM16, a master regulator of BAT differentiation, and enhances the stability of PRDM16 protein. These results define Tyk2 and Stat3 as critical determinants of brown fat lineage and suggest that altered levels of Tyk2 are associated with obesity in both rodents and humans.

Leptin resistance in a polygenic, hyperleptinemic animal model of obesity and NIDDM : Psammomys obesus

OBJECTIVE: To investigate the effects of leptin administration to Psammomys obesus, a polygenic animal model of obesity and type 2 diabetes mellitus.

DESIGN: Longitudinal intervention study utilising three separate leptin treatment protocols lasting 7-14 d.

MEASUREMENTS: Body weight and food intake were measured daily, body fat and muscle content were estimated by carcass analysis on completion of the study. Blood glucose, plasma insulin, leptin, triglycerides and cholesterol were measured at baseline and twice each week during the study.

RESULTS: Relatively high doses of leptin were required to significantly reduce food intake and body fat content in lean Psammomys obesus, but had no discernible effect on their obese littermates.

CONCLUSION: As a species, Psammomys obesus appear to be relatively insensitive to the effects of leptin administration, compared with other rodents. Obese Psammomys obesus are leptin resistant relative to their lean littermates.

Are voluntary wheel running and open-field behavior correlated in mice? Different answers from comparative and artificial selection approaches

Voluntary wheel running and open-field behavior are probably the two most widely used measures of locomotion in laboratory rodents. We tested whether these two behaviors are correlated in mice using two approaches: the phylogenetic comparative method using inbred strains of mice and an ongoing artificial selection experiment on voluntary wheel running. After taking into account the measurement error and phylogenetic relationships among inbred strains, we obtained a significant positive correlation between distance run on wheels and distance moved in the open-field for both sexes. Thigmotaxis was negatively correlated with distance run on wheels in females but not in males. By contrast, mice from four replicate lines bred for high wheel running did not differ in either distance covered or thigmotaxis in the open field as compared with mice from four non-selected control lines. Overall, results obtained in the selection experiment were generally opposite to those observed among inbred strains. Possible reasons for this discrepancy are discussed.

Overexpression of carnitine palmitoyltransferase I in skeletal muscle in vivo increases fatty acid oxidation and reduces triacylglycerol esterification

A key regulatory point in the control of fatty acid (FA) oxidation is thought to be transport of FAs across the mitochondrial membrane by carnitine palmitoyltransferase I (CPT I). To investigate the role of CPT I in FA metabolism, we used in vivo electrotransfer (IVE) to locally overexpress CPT I in muscle of rodents. A vector expressing the human muscle isoform of CPT I was electrotransferred into the right lateral muscles of the distal hindlimb [tibialis cranialis (TC) and extensor digitorum longus (EDL)] of rats, and a control vector expressing GFP was electrotransferred into the left muscles. Initial studies showed that CPT I protein expression peaked 7 days after IVE (+104%, P < 0.01). This was associated with an increase in maximal CPT I activity (+30%, P < 0.001) and a similar increase in palmitoyl-CoA oxidation (+24%; P < 0.001) in isolated mitochondria from the TC. Importantly, oxidation of the medium-chain FA octanoyl-CoA and CPT I sensitivity to inhibition by malonyl-CoA were not altered by CPT I overexpression. FA oxidation in isolated EDL muscle strips was increased with CPT I overexpression (+28%, P < 0.01), whereas FA incorporation into the muscle triacylglycerol (TAG) pool was reduced (−17%, P < 0.01). As a result, intramyocellular TAG content was decreased with CPT I overexpression in both the TC (−25%, P < 0.05) and the EDL (−45%, P < 0.05). These studies demonstrate that acute overexpression of CPT I in muscle leads to a repartitioning of FAs away from esterification and toward oxidation and highlight the importance of CPT I in regulating muscle FA metabolism.

Oral treatment with Cu(II)(atsm) increases mutant SOD1 in vivo but protects motor neurons and improves the phenotype of a transgenic mouse model of amyotrophic lateral sclerosis

Mutations in the metallo-protein Cu/Zn-superoxide dismutase (SOD1) cause amyotrophic lateral sclerosis (ALS) in humans and an expression level-dependent phenotype in transgenic rodents. We show that oral treatment with the therapeutic agent diacetyl-bis(4-methylthiosemicarbazonato)copper(II) [Cu(II)(atsm)] increased the concentration of mutant SOD1 (SOD1G37R) in ALS model mice, but paradoxically improved locomotor function and survival of the mice. To determine why the mice with increased levels of mutant SOD1 had an improved phenotype, we analyzed tissues by mass spectrometry. These analyses revealed most SOD1 in the spinal cord tissue of the SOD1G37R mice was Cu deficient. Treating with Cu(II)(atsm) decreased the pool of Cu-deficient SOD1 and increased the pool of fully metallated (holo) SOD1. Tracking isotopically enriched (65)Cu(II)(atsm) confirmed the increase in holo-SOD1 involved transfer of Cu from Cu(II)(atsm) to SOD1, suggesting the improved locomotor function and survival of the Cu(II)(atsm)-treated SOD1G37R mice involved, at least in part, the ability of the compound to improve the Cu content of the mutant SOD1. This was supported by improved survival of SOD1G37R mice that expressed the human gene for the Cu uptake protein CTR1. Improving the metal content of mutant SOD1 in vivo with Cu(II)(atsm) did not decrease levels of misfolded SOD1. These outcomes indicate the metal content of SOD1 may be a greater determinant of the toxicity of the protein in mutant SOD1-associated forms of ALS than the mutations themselves. Improving the metal content of SOD1 therefore represents a valid therapeutic strategy for treating ALS caused by SOD1.

Snakes on an island: independent introductions have different potentials for invasion

Snakes introduced to islands can be devastating to naïve native fauna. However, introduced populations must establish before range expansion (invasion) can occur. The factors that can determine successful invasion are those associated with the introduction event (e.g., characteristics of the founding population), the location (e.g., suitable environment and prey availability) and the species (e.g. life history characteristics). Here, we collected morphometric, ecological and genetic data on the recently introduced California Kingsnake (Lampropeltis californiae) in Gran Canaria. We found that snakes occurring at two locations a few 10 s of km apart do not represent the same population. Genetic analyses confirmed significant genetic difference (FST = 0.184; Dest = 0.341), and that despite being inbred (Fis = 0.245–0.257) the populations had high levels of diversity (Ho = 0.485–0.490; allelic richness = 4.875–6.364). Snakes at the different Gran Canaria locations were significantly different in morphology (colouration, mass, length and age), fitness (egg production) and diet (rodents, skinks, lizards and geckos), supporting a hypothesis of separate founding groups in combination with local environmental heterogeneity leading to variation between these populations. We concluded that one population was more successful than the other in reproduction and recruitment, and may be having a greater impact on endemic reptiles. We recommend greater eradication effort for this population, as well as monitoring of local fauna at all locations to access the impact of predation.

Metabolic rate and thermal conductance of lemmings from high-arctic Canada and Siberia

The arctic climate places high demands on the energy metabolism of its inhabitants. We measured resting (RMR) and basal metabolic rates (BMR), body temperatures, and dry and wet thermal conductances in summer morphs of the lemmings Dicrostonyx groenlandicus and Lemmus trimucronatus in arctic Canada, and the BMR of D. torquatus, D. groenlandicus, L. sibiricus, L. bungei and L. trimucronatus in Siberia. In contrast to previous studies the data were collected on animals that had spent only a limited time in captivity. All parameters were analysed in relation to the variations in body mass (20-90 g). Body temperature and BMR were lower in D. groenlandicus than L. trimucronatus, which coincides with greater longevity in the former species. Wet and dry thermal conductances of both species were similar and comparable with those of other Myomorpha (mouse-type rodents), indicating no evidence for a previously claimed lower thermal conductance in lemmings. BMR in lemmings appeared to be higher than in other Arvicolidae (voles, lemmings and muskrats), which could relate to their typically high-latitude distribution. However, the more southerly living Lemmus species had higher BMR than the more northerly living Dicrostonyx species, which may be explained by the former having a relatively low-quality diet.

Energy expenditure and personality in wild chipmunks

© 2015, Springer-Verlag Berlin Heidelberg. According to the “pace-of-life syndrome” concept, slow-fast life-history strategies favored under different ecological conditions should lead to co-adaptations between metabolic rate and personality traits such as activity, exploration, and boldness. Although the relationships between resting metabolic rate (RMR) and personality traits have been recently tested several times, we still do not know whether personality is related to the daily energy expenditure (DEE) of free-living individuals in their natural habitat. The objectives of this study were to assess the links between RMR, DEE, and two personality traits (exploration in an open-field and docility during handling) in wild eastern chipmunks (Tamias striatus). Using a multivariate mixed model, we found that exploration and docility were significantly correlated at the among-individual level, confirming the presence of a behavioral syndrome within our population. We also found that exploration, but not docility, was negatively correlated with DEE. Hence, fast explorers show lower DEE levels than slow explorers, independently of RMR and docility. This result adds to an increasingly large (and complex) literature reporting the impacts of personality traits on the biology, ecology, and physiology of animals in their natural environment.

Ascorbic acid supplementation improves skeletal muscle oxidative stress and insulin sensitivity in people with type 2 diabetes: findings of a randomized controlled study

AIM/HYPOTHESIS: Skeletal muscle insulin resistance and oxidative stress are characteristic metabolic disturbances in people with type 2 diabetes. Studies in insulin resistant rodents show an improvement in skeletal muscle insulin sensitivity and oxidative stress following antioxidant supplementation. We therefore investigated the potential ameliorative effects of antioxidant ascorbic acid (AA) supplementation on skeletal muscle insulin sensitivity and oxidative stress in people with type 2 diabetes. METHODS: Participants with stable glucose control commenced a randomized cross-over study involving four months of AA (2×500mg/day) or placebo supplementation. Insulin sensitivity was assessed using a hyperinsulinaemic, euglycaemic clamp coupled with infusion of 6,6-D2 glucose. Muscle biopsies were measured for AA concentration and oxidative stress markers that included basal measures (2',7'-dichlorofluorescin [DCFH] oxidation, ratio of reduced-to-oxidized glutathione [GSH/GSSG] and F2-Isoprostanes) and insulin-stimulated measures (DCFH oxidation). Antioxidant concentrations, citrate synthase activity and protein abundances of sodium-dependent vitamin C transporter 2 (SVCT2), total Akt and phosphorylated Akt (ser473) were also measured in muscle samples. RESULTS: AA supplementation significantly increased insulin-mediated glucose disposal (delta rate of glucose disappearance; ∆Rd) (p=0.009), peripheral insulin-sensitivity index (p=0.046), skeletal muscle AA concentration (p=0.017) and muscle SVCT2 protein expression (p=0.008); but significantly decreased skeletal muscle DCFH oxidation during hyperinsulinaemia (p=0.007) when compared with placebo. Total superoxide dismutase activity was also lower following AA supplementation when compared with placebo (p=0.006). Basal oxidative stress markers, citrate synthase activity, endogenous glucose production, HbA1C and muscle Akt expression were not significantly altered by AA supplementation. CONCLUSIONS/INTERPRETATION: In summary, oral AA supplementation ameliorates skeletal muscle oxidative stress during hyperinsulinaemia and improves insulin-mediated glucose disposal in people with type 2 diabetes. Findings implicate AA supplementation as a potentially inexpensive, convenient, and effective adjunct therapy in the treatment of insulin resistance in people with type 2 diabetes.

A continental-scale analysis of feral cat diet in Australia

Aim Reducing the impacts of feral cats (Felis catus) is a priority for conservation managers across the globe, and success in achieving this aim requires a detailed understanding of the species’ ecology across a broad spectrum of climatic and environmental conditions. We reviewed the diet of the feral cat across Australia and on Australian territorial islands, seeking to identify biogeographical patterns in dietary composition and diversity, and use the results to consider how feral cats may best be managed. Location Australia and its territorial islands. Methods Using 49 published and unpublished data sets, we modelled trophic diversity and the consumption of eight food groups against latitude, longitude, mean temperature, precipitation, environmental productivity and climate-habitat regions. Results We recorded 400 vertebrate species that feral cats feed on or kill in Australia, including 28 IUCN Red List species. We found evidence of continental- scale prey-switching from rabbits to small mammals, previously recorded only at the local scale. The consumption of arthropods, reptiles, rabbits, rodents and medium-sized native mammals varied with different combinations of latitude, longitude, mean annual precipitation, temperature and environmental productivity. The frequency of rodents and dasyurids in cats’ diets increased as rabbit consumption decreased. Main conclusions The feral cat is an opportunistic, generalist carnivore that consumes a diverse suite of vertebrate prey across Australia. It uses a facultative feeding strategy, feeding mainly on rabbits when they are available, but switching to other food groups when they are not. Control programmes aimed at culling rabbits could potentially decrease the availability of a preferred food source for cats and then lead to greater predation pressure on native mammals. The interplay between cat diet and prey species diversity at a continental scale is complex, and thus cat management is likely to be necessary and most effective at the local landscape level.

A game of cat-and-mouse: microhabitat influences rodent foraging in recently burnt but not long unburnt shrublands

We investigated the influence of vegetation structure and fire history on the foraging behavior of small rodents (Notomys mitchellii, Pseudomys hermannsburgensis, and Mus musculus) by conducting giving-up density (GUD) experiments in recently burnt (9–13 years since last fire) and long unburnt shrublands (> 40 years), and open and sheltered microhabitats, in a semiarid region of Western Australia. We predicted that rodents would spend less time foraging in recently burnt shrublands and open microhabitat and that the influence of microhabitat would be weaker in long unburnt compared to more recently burnt vegetation. Our findings show that fire history and microhabitat structure influence the foraging behavior of the study species and that the influence of microhabitat varies between fire histories. GUDs were higher in long unburnt vegetation and in open microhabitats. There was a microhabitat effect in recently burnt vegetation, but not in long unburnt. Rodents foraged more in sheltered microhabitats probably because predator encounters are less likely to occur there and it provides them with greater refuge from predation. The presence of a microhabitat effect in recently burnt, but not long unburnt vegetation suggests that understory vegetation density is more important in mediating predation risk than canopy density. Future studies of small mammal responses to land management actions should include behavioral, as well as population-level responses to differing fire regimes.

Invasive predators and global biodiversity loss.

Invasive species threaten biodiversity globally, and invasive mammalian predators are particularly damaging, having contributed to considerable species decline and extinction. We provide a global metaanalysis of these impacts and reveal their full extent. Invasive predators are implicated in 87 bird, 45 mammal, and 10 reptile species extinctions-58% of these groups' contemporary extinctions worldwide. These figures are likely underestimated because 23 critically endangered species that we assessed are classed as "possibly extinct." Invasive mammalian predators endanger a further 596 species at risk of extinction, with cats, rodents, dogs, and pigs threatening the most species overall. Species most at risk from predators have high evolutionary distinctiveness and inhabit insular environments. Invasive mammalian predators are therefore important drivers of irreversible loss of phylogenetic diversity worldwide. That most impacted species are insular indicates that management of invasive predators on islands should be a global conservation priority. Understanding and mitigating the impact of invasive mammalian predators is essential for reducing the rate of global biodiversity loss.