Hypoxia exposure blunts angiogenic signaling and upregulates the antioxidant system in endothelial cells derived from elephant seals.

BACKGROUND: Elephant seals exhibit extreme hypoxemic tolerance derived from repetitive hypoxia/reoxygenation episodes they experience during diving bouts. Real-time assessment of the molecular changes underlying protection against hypoxic injury in seals remains restricted by their at-sea inaccessibility. Hence, we developed a proliferative arterial endothelial cell culture model from elephant seals and used RNA-seq, functional assays, and confocal microscopy to assess the molecular response to prolonged hypoxia. RESULTS: Seal and human endothelial cells exposed to 1% O2 for up to 6 h respond differently to acute and prolonged hypoxia. Seal cells decouple stabilization of the hypoxia-sensitive transcriptional regulator HIF-1α from angiogenic signaling. Rapid upregulation of genes involved in glutathione (GSH) metabolism supports the maintenance of GSH pools, and intracellular succinate increases in seal but not human cells. High maximal and spare respiratory capacity in seal cells after hypoxia exposure occurs in concert with increasing mitochondrial branch length and independent from major changes in extracellular acidification rate, suggesting that seal cells recover oxidative metabolism without significant glycolytic dependency after hypoxia exposure. CONCLUSIONS: We found that the glutathione antioxidant system is upregulated in seal endothelial cells during hypoxia, while this system remains static in comparable human cells. Furthermore, we found that in contrast to human cells, hypoxia exposure rapidly activates HIF-1 in seal cells, but this response is decoupled from the canonical angiogenesis pathway. These results highlight the unique mechanisms that confer extraordinary tolerance to limited oxygen availability in a champion diving mammal.

Circadian coupling of mitochondria in a deep-diving mammal.

Regulation of mitochondrial oxidative phosphorylation is essential to match energy supply to changing cellular energy demands, and to cope with periods of hypoxia. Recent work implicates the circadian molecular clock in control of mitochondrial function and hypoxia sensing. Because diving mammals experience intermittent episodes of severe hypoxia, with diel patterning in dive depth and duration, it is interesting to consider circadian-mitochondrial interaction in this group. Here, we demonstrate that the hooded seal (Cystophora cristata), a deep-diving Arctic pinniped, shows strong daily patterning of diving behaviour in the wild. Cultures of hooded seal skin fibroblasts exhibit robust circadian oscillation of the core clock genes per2 and arntl. In liver tissue collected from captive hooded seals, expression of arntl was some 4-fold higher in the middle of the night than in the middle of the day. To explore the clock-mitochondria relationship, we measured the mitochondrial oxygen consumption in synchronized hooded seal skin fibroblasts and found a circadian variation in mitochondrial activity, with higher coupling efficiency of complex I coinciding with the trough of arntl expression. These results open the way for further studies of circadian-hypoxia interactions in pinnipeds during diving.

The roles of brain lipids and polar metabolites in the hypoxia tolerance of deep-diving pinnipeds.

Lipids make up more than half of the human brain's dry weight, yet the composition and function of the brain lipidome is not well characterized. Lipids not only provide the structural basis of cell membranes, but also take part in a wide variety of biochemical processes. In neurodegenerative diseases, lipids can facilitate neuroprotection and serve as diagnostic biomarkers. The study of organisms adapted to extreme environments may prove particularly valuable in understanding mechanisms that protect against stressful conditions and prevent neurodegeneration. The brain of the hooded seal (Cystophora cristata) exhibits a remarkable tolerance to low tissue oxygen levels (hypoxia). While neurons of most terrestrial mammals suffer irreversible damage after only short periods of hypoxia, in vitro experiments show that neurons of the hooded seal display prolonged functional integrity even in severe hypoxia. How the brain lipidome contributes to the hypoxia tolerance of marine mammals has been poorly studied. We performed an untargeted lipidomics analysis, which revealed that lipid species are significantly modulated in marine mammals compared with non-diving mammals. Increased levels of sphingomyelin species may have important implications for efficient signal transduction in the seal brain. Substrate assays also revealed elevated normoxic tissue levels of glucose and lactate, which suggests an enhanced glycolytic capacity. Additionally, concentrations of the neurotransmitters glutamate and glutamine were decreased, which may indicate reduced excitatory synaptic signaling in marine mammals. Analysis of hypoxia-exposed brain tissue suggests that these represent constitutive mechanisms rather than an induced response towards hypoxic conditions.

Mercury Bioaccumulation and Cortisol Interact to Influence Endocrine and Immune Biomarkers in a Free-Ranging Marine Mammal.

Mercury bioaccumulation from deep-ocean prey and the extreme life history strategies of adult female northern elephant seals (Mirounga angustirostris) provide a unique system to assess the interactive effects of mercury and stress on animal health by quantifying blood biomarkers in relation to mercury (skeletal muscle and blood mercury) and cortisol concentrations. The thyroid hormone thyroxine (tT4) and the antibody immunoglobulin E (IgE) were associated with mercury and cortisol concentrations interactively, where the magnitude and direction of the association of each biomarker with mercury or cortisol changed depending on the concentration of the other factor. For example, when cortisol concentrations were lowest, tT4 was positively related to muscle mercury, whereas tT4 had a negative relationship with muscle mercury in seals that had the highest cortisol concentrations. Additionally, we observed that two thyroid hormones, triiodothyronine (tT3) and reverse triiodothyronine (rT3), were negatively (tT3) and positively (rT3) associated with mercury concentrations and cortisol in an additive manner. As an example, tT3 concentrations in late breeding seals at the median cortisol concentration decreased by 14% across the range of observed muscle mercury concentrations. We also observed that immunoglobulin M (IgM), the pro-inflammatory cytokine IL-6 (IL-6), and a reproductive hormone, estradiol, were negatively related to muscle mercury concentrations but were not related to cortisol. Specifically, estradiol concentrations in late molting seals decreased by 50% across the range of muscle mercury concentrations. These results indicate important physiological effects of mercury on free-ranging apex marine predators and interactions between mercury bioaccumulation and extrinsic stressors. Deleterious effects on animals' abilities to maintain homeostasis (thyroid hormones), fight off pathogens and disease (innate and adaptive immune system), and successfully reproduce (endocrine system) can have significant individual- and population-level consequences.

I've got you under my skin: inflammatory response to elephant seal's lice.

Seals (Phocidae) undergo an annual cycle of moulting that implies hair regeneration, and in the case of southern elephant seals, it also involves the superficial strata of the epidermis. Therefore, surviving the moulting period is crucial for their obligate and permanent ectoparasites. Throughout evolutionary time, sucking lice (Echinophtiriidae) have developed morphological, behavioural and ecological adaptations to cope with the amphibious lifestyle of their hosts. Lepidophthirus macrorhini, the Southern elephant seal louse species, faces the additional challenge of surviving attached to the host during the moulting period. Since lice live on the skin, L. macrorhini has developed a unique survival strategy by piercing the skin of their host, thus keeping them protected from moulting. During fieldwork in Patagonia and Antarctica, skin samples with lice within were collected for histological analysis to assess whether these parasites caused damage to the host. Lice generate an inflammatory process in the host's dermis, and these lesions could alter the normal chemical and mechanical protective properties of the skin facilitating secondary infections. Further studies that analyse the potential pathogens in those skin lesions are necessary to properly assess the real impact of ectoparasites on their host health.

Male grey seal commits fatal sexual interaction with adult female harbour seals in the German Wadden Sea.

Males of several seal species are known to show aggressive copulating behaviour, which can lead to injuries to or suffocation of females. In the North Sea, grey seal predation on harbour seals including sexual harassment is documented and represents violent interspecific interaction. In this case series, we report pathological and molecular/genetic findings of 11 adult female harbour seals which were found dead in Schleswig-Holstein, Germany, within 41 days. Several organs of all animals showed haemorrhages and high loads of bacteria, indicating their septic spread. All females were pregnant or had recently been pregnant. Abortion was confirmed in three cases. Lacerations were seen in the uterus and vagina in six cases, in which histology of three individuals revealed severe suppurative inflammation with intralesional spermatozoa. Molecular analysis of vaginal swabs and paraffin-embedded samples of the vagina identified grey seal DNA, suggesting violent interspecific sexual interaction with fatal outcome due to septicaemia. This is the first report of female harbour seals dying after coercive copulation by a male grey seal in the Wadden Sea.

Antioxidant response to cadmium exposure in primary skeletal muscle cells isolated from humans and elephant seals.

Cadmium (Cd) occurs naturally; however, its concentration can increase with anthropogenic activities. Excess Cd increases reactive oxygen species (ROS) production and oxidative damage, which can lead to pathological conditions. Marine mammals accumulate Cd in the liver and the kidney; yet, there are no reports of Cd-associated tissue damage in whales, seals or dolphins. Response to Cd exposure (0-5.0 µM CdCl2 for 1-12 h) was analyzed and compared in primary skeletal muscle cells isolated from northern elephant seals (Mirounga angustirostris) and humans (Homo sapiens). Antioxidant enzyme activities (glutathione S-transferase, glutathione reductase, glutathione peroxidase), glutathione concentration, and protein carbonyl levels (an indicator of oxidative damage) were quantified. Glutathione levels were higher in northern elephant seal than in human cells. Protein carbonyl content in cells exposed to Cd was lower and had a smaller variability range in elephant seals than in humans. Generalized linear models (GLIM) identified Cd exposure and antioxidant defenses as significant contributors to protein carbonyl variability in human but not in elephant seal cells. These results suggest that the previously observed differences in circulating and tissue glutathione levels between marine and terrestrial mammals are maintained under cell culture conditions and that northern elephant seal and human muscle cells respond differently to Cd exposure. The results also suggest that the observed differences could potentially be associated with the protective mechanisms that allow northern elephant seals to tolerate extreme conditions that result in increased ROS generation (e.g. diving, sleep apnea, fasting) with no oxidative damage.

Hooded seal Cystophora cristata foraging areas in the Northeast Atlantic Ocean-Investigated using three complementary methods.

Identifying environmental characteristics that define the ecological niche of a species is essential to understanding how changes in physical conditions might affect its distribution and other aspects of its ecology. The present study used satellite relay data loggers (SRDLs) to study habitat use by Northeast Atlantic hooded seals (N = 20; 9 adult females, 3 adult males, and 8 juveniles). Three different methods were used in combination to achieve maximum insight regarding key foraging areas for hooded seals in this region, which have decline by 85% in recent decades: 1) first passage time (FPT); 2) vertical transit rate and; 3) change in dive drift rate. Generalized additive mixed models (GAMM) were applied to each method to determine whether specific habitat characteristics were associated with foraging. Separate models were run for the post-molting and the post-breeding seasons; sex and age classes were included in the GAMMs. All three methods highlighted a few common geographic areas as being important foraging zones; however, there were also some different areas identified by the different methods, which highlights the importance of using multiple indexes when analyzing tracking and diving data to study foraging behavior. Foraging occurred most commonly in relatively shallow areas with high Sea Surface Temperatures (SST), corresponding to continental shelf areas with Atlantic Water masses. All age and sex classes overlapped spatially to some extent, but the different age and sex groups showed differences in the bathymetry of their foraging areas as well as in their vertical use of the water column. When foraging, pups dove in the upper part of the water column in relatively deep areas. Adult females foraged relatively shallowly in deep water areas too, though in shallower areas than pups. Adult males foraged close to the bottom in shallower areas.

Carbon, nitrogen and sulphur isotopic fractionation in captive juvenile hooded seal (Cystophora cristata): Application for diet analysis.

RATIONALE: Intrinsic biogeochemical markers, such as stable isotope ratios of carbon, nitrogen and sulphur, are increasingly used to trace the trophic ecology of marine top predators. However, insufficient knowledge of fractionation processes in tissues continues to hamper the use of these markers. METHODS: We performed a controlled feeding experiment with eight juvenile hooded seals (Cystophora cristata) that were held on a herring-based diet (Clupea harengus) for two years. Stable isotope ratios were measured via isotope ratio mass spectrometry in three of their tissues and related to values of these markers in their diet. RESULTS: Diet-tissue isotope enrichment (trophic enrichment factor, TEF) values between dietary herring and seal tissues for carbon (Δ13 C) were +0.7 ‰ for red blood cells, +1.9 ‰ for hair and +1.1 ‰ for muscle. The TEFs for nitrogen trophic (Δ15 N) were +3.3 ‰ for red blood cells, +3.6 ‰ for hair and +4.3 ‰ for muscle. For sulphur, the Δ34 S values were +1.1 ‰ for red blood cells, +1.0 ‰ for hair and +0.9 ‰ for muscle. CONCLUSIONS: These enrichment values were greater than those previously measured in adult seals. This increase may be related to the higher rate of protein synthesis and catabolism in growing animals. This study is the first report on sulphur isotope enrichment values for a marine mammal species.

Hooded seal (Cystophora cristata) pups ingest snow and seawater during their post-weaning fast.

The purpose of this study was to evaluate the importance of exogenous water intake (snow/seawater) in hooded seal (Cystophora cristata) pups during their post-weaning fast. In this study, five hooded seal pups had ad lib access to snow and seawater for the first 12 and last 21 days of their post-weaning fast, respectively. Total body water and water flux were determined during both exposure periods by use of the tritiated water method. Blood samples were collected to monitor changes in hematocrit, plasma urea and plasma osmolality. Body mass loss was on average 0.36 kg day-1. Average total body water changed from 15.7 to 11.4 L, while total water influx changed from 15 to 18 mL day-1 kg-1 during snow and seawater exposure, respectively. Of this influx an average of 35% can be attributed to metabolic water, while approximately 8% was due to respiratory water influx. Interestingly, 56 and 58% of the total water influx was due to snow and seawater ingestion, respectively, amounting to 8 mL day-1 kg-1 snow (counted as liquid water) and 10 mL day-1 kg-1 seawater. Based on the results of the plasma parameters it is concluded that fasting hooded seal pups maintain water balance and homeostasis when access to snow or seawater is permitted. It is further concluded that snow and seawater intake, in addition to metabolic and respiratory water, is important for maintenance of water balance and excretion of urea during the post-weaning fast of hooded seal pups.

Synaptic transmission despite severe hypoxia in hippocampal slices of the deep-diving hooded seal.

Brain neurons of the deep-diving hooded seal (Cystophora cristata) are known to be inherently hypoxia tolerant. Here, we have used in vitro field potential recordings in hippocampal slices to compare effects of severe hypoxia on synaptic transmission in hooded seals vs. non-diving mammals. Synaptic responses of mice (Mus musculus) to hypoxia were in accordance with previously published data. Hippocampal slices of reindeer (Rangifer tarandus), an alternative large-mammal non-diving model, behaved in a similar way as mouse slices, in that synaptic activity disappeared rapidly without recovery after >20min in hypoxia. The synaptic activity of hooded seal slices decreased in hypoxia, but unlike mice and reindeer, it remained at >30% of the normoxic amplitude throughout 3h of severe hypoxia. Also, upon reoxygenation, the signal recovered to ∼50% of the pre-challenge (normoxic) amplitude. The AMPA-type glutamate receptor antagonist CNQX eliminated this signal, showing that it was not an artifact. Paired pulse facilitation (PPF), typically associated with increased presynaptic calcium (Ca2+) levels, was significantly reduced in the seal slices. We propose that the build-up of Ca2+ concentration is limited in seal presynaptic terminals, possibly due to a high Ca2+ buffering capacity, which could explain both the attenuated PPF and the remarkable neural hypoxia tolerance of this species. Although we found no significant hypoxia-induced upregulation of mRNA for the Ca2+ binding proteins calbindin d28k or parvalbumin in hooded seal hippocampal slices, a recent study reports very high transcript levels of the Ca2+ binding protein S100B in this species, which is in support of the hypothesis.

Meals on Wheels? A Decade of Megafaunal Visual and Acoustic Observations from Offshore Oil & Gas Rigs and Platforms in the North and Irish Seas.

A decade of visual and acoustic detections of marine megafauna around offshore Oil & Gas (O&G) installations in the North and Irish Seas are presented. Marine megafauna activity was monitored visually and acoustically by Joint Nature Conservation Committee (JNCC) qualified and experienced Marine Mammal Observers (MMO) and Passive Acoustic Monitoring (PAM) Operators respectively, with real-time towed PAM in combination with industry standard software, PAMGuard. Monitoring was performed during routine O&G industrial operations for underwater noise mitigation purposes, and to ensure adherence to regulatory guidelines. Incidental sightings by off-effort MMOs and installation crew were also reported. Visual and acoustic monitoring spanned 55 non-consecutive days between 2004 and 2014. A total of 47 marine mammal sightings were recorded by MMOs on dedicated watch, and 10 incidental sightings of marine megafauna were reported over 10 years. Species included: harbour porpoise (Phocoena phocoena), Atlantic white-sided dolphin (Lagenorhynchus acutus), white beaked dolphin (Lagenorhynchus albirostris), common dolphin (Delphinus delphis), minke whale (Balaenoptera acutorostrata), common seal (Phoca vitulina), grey seal (Halichoerus grypus) and, basking shark (Cetorhinus maximus). Passive Acoustic Monitoring was conducted on two occasions in 2014; 160 PAM hours over 12 days recorded a total of 308 individual clicks identified as harbour porpoises. These appear to be the first such acoustic detections obtained from a North Sea drilling rig whilst using a typically configured hydrophone array designed for towing in combination with real-time PAMGuard software. This study provides evidence that marine megafauna are present around mobile and stationary offshore O&G installations during routine operational activities. On this basis, Environmental Impact Assessments (EIAs) for decommissioning O&G platforms should be carried-out on a case-by-case basis, and must include provisions for hitherto overlooked marine megafauna.

An atypical distribution of lactate dehydrogenase isoenzymes in the hooded seal (Cystophora cristata) brain may reflect a biochemical adaptation to diving.

The brains of some diving mammals can withstand periods of severe hypoxia without signs of deleterious effects. This may in part be due to an enhanced cerebral capacity for anaerobic energy production. Here, we have tested this hypothesis by comparing various parameters of the lactate dehydrogenase (LDH) in the brain of the hooded seal (Cystophora cristata) with those in the brains of the ferret (Mustela putorius furo) and mouse (Mus musculus). We found that mRNA and protein expression of lactate dehydrogenase a (LDHA) and lactate dehydrogenase b (LDHB), and also the LDH activity were significantly higher in the ferret brain than in brains of the hooded seal and the mouse (p < 0.0001). No conspicuous differences in the LDHA and LDHB sequences were observed. There was also no difference in the buffering capacities of the brains. Thus, an enhanced capacity for anaerobic energy production likely does not explain the higher hypoxia tolerance of the seal brain. However, the brain of the hooded seal had higher relative levels of LDHB isoenzymes (LDH1 and LDH2) compared to the non-diving mammals. Moreover, immunofluorescence studies showed more pronounced co-localization of LDHB and glial fibrillary acidic protein in the cortex of the hooded seal. Since LDHB isoenzymes primarily catalyze the conversion of lactate to pyruvate, this finding suggests that the contribution of astrocytes to the brain aerobic metabolism is higher in the hooded seal than in non-diving species. The cerebral tolerance of the hooded seal to hypoxia may therefore partly rely on different LDH isoenzymes distribution.

The better to eat you with: the comparative feeding morphology of phocid seals (Pinnipedia, Phocidae).

One adaptation crucial to the survival of mammalian lineages that secondarily transitioned from land to water environments was the ability to capture and consume prey underwater. Phocid seals have evolved diverse feeding strategies to feed in the marine environment, and the objectives of this study were to document the specialized feeding morphologies and identify feeding strategies used by extant phocids. This study used principal component analysis (PCA) to determine the major axes of diversification in the skull for all extant phocid taxa and the recently extinct Caribbean monk seal (n = 19). Prey data gathered from the literature and musculoskeletal data from dissections were included to provide a comprehensive description of each feeding strategy. Random Forest analysis was used to determine the morphological, ecological and phylogenetic variables that best described each feeding strategy. There is morphological evidence for four feeding strategies in phocids: filter; grip and tear; suction; and pierce feeding. These feeding strategies are supported by quantitative cranial and mandibular characters, dietary information, musculoskeletal data and, for some species, behavioral observations. Most phocid species are pierce feeders, using a combination of biting and suction to opportunistically catch prey. Grip and tear and filter feeding are specialized strategies with specific morphological adaptations. These unique adaptations have allowed leopard seals (Hydrurga leptonyx) and crabeater seals (Lobodon carcinophaga) to exploit novel ecological niches and prey types. This study provides the first cranial and mandibular morphological evidence for the use of specialized suction feeding in hooded seals (Cystophora cristata), northern elephant seals (Mirounga angustirostris) and southern elephant seals (Mirounga leonina). The most important variables in determining the feeding strategy of a given phocid species were cranial and mandibular shape, diet, and phylogeny. These results provide a framework for understanding the evolution and adaptability of feeding strategies employed by extant phocid species, and these findings can be applied to other pinniped lineages and extinct taxa.

To each its own: Thermoregulatory strategy varies among neonatal polar phocids.

Cold environmental conditions and small body size promote heat loss and may create thermoregulatory challenges for marine mammals born in polar regions. However, among polar-born phocid seal species there are variations in physical attributes and environmental conditions at birth, allowing for an interesting contrast in thermoregulatory strategy. We compared thermoregulatory strategies through morphometrics, sculp attributes (conductivity and resistance), nonshivering thermogenesis (NST via uncoupling protein 1; UCP1), and muscle thermogenesis (via enzyme activity) in neonatal harp (Pagophilus groenlandicus), hooded (Cystophora cristata), and Weddell seals (Leptonychotes weddellii). Harp seals are the smallest at birth (9.8±0.7 kg), rely on lanugo (82.49±3.70% of thermal resistance), and are capable of NST through expression of UCP1 in brown adipose tissue (BAT). In contrast, hooded seal neonates (26.8±1.3 kg) have 2.06±0.23 cm of blubber, accounting for 38.19±6.07% of their thermal resistance. They are not capable of NST, as UCP1 is not expressed. The large Weddell seal neonates (31.5±4.9 kg) rely on lanugo (89.85±1.25% of thermal resistance) like harp seals, but no evidence of BAT was found. Muscle enzyme activity was highest in Weddell seal neonates, suggesting that they rely primarily on muscle thermogenesis. Similar total thermal resistance, combined with marked differences in thermogenic capacity of NST and ST among species, strongly supports that thermoregulatory strategy in neonatal phocids is more closely tied to pups' surface area to volume ratio (SA:V) and potential for early water immersion rather than mass and ambient environmental conditions.

Investigating annual diving behaviour by hooded seals (Cystophora cristata) within the Northwest Atlantic Ocean.

With the exception of relatively brief periods when they reproduce and moult, hooded seals, Cystophora cristata, spend most of the year in the open ocean where they undergo feeding migrations to either recover or prepare for the next fasting period. Valuable insights into habitat use and diving behaviour during these periods have been obtained by attaching Satellite Relay Data Loggers (SRDLs) to 51 Northwest (NW) Atlantic hooded seals (33 females and 18 males) during ice-bound fasting periods (2004-2008). Using General Additive Models (GAMs) we describe habitat use in terms of First Passage Time (FPT) and analyse how bathymetry, seasonality and FPT influence the hooded seals' diving behaviour described by maximum dive depth, dive duration and surface duration. Adult NW Atlantic hooded seals exhibit a change in diving activity in areas where they spend >20 h by increasing maximum dive depth, dive duration and surface duration, indicating a restricted search behaviour. We found that male and female hooded seals are spatially segregated and that diving behaviour varies between sexes in relation to habitat properties and seasonality. Migration periods are described by increased dive duration for both sexes with a peak in May, October and January. Males demonstrated an increase in dive depth and dive duration towards May (post-breeding/pre-moult) and August-October (post-moult/pre-breeding) but did not show any pronounced increase in surface duration. Females dived deepest and had the highest surface duration between December and January (post-moult/pre-breeding). Our results suggest that the smaller females may have a greater need to recover from dives than that of the larger males. Horizontal segregation could have evolved as a result of a resource partitioning strategy to avoid sexual competition or that the energy requirements of males and females are different due to different energy expenditure during fasting periods.

Activation of systemic, but not local, renin-angiotensin system is associated with upregulation of TNF-α during prolonged fasting in northern elephant seal pups.

Northern elephant seal pups naturally endure a 2-3 month post-weaning fast that is associated with activation of systemic renin-angiotensin system (RAS), a decrease in plasma adiponectin (Acrp30), and insulin resistance (IR)-like conditions. Angiotensin II (Ang II) and tumor necrosis factor-alpha (TNF-α) are potential causal factors of IR, while Acrp30 may improve insulin signaling. However, the effects of fasting-induced activation of RAS on IR-like conditions in seals are not well described. To assess the effects of prolonged food deprivation on systemic and local RAS, and their potential contribution to TNF-α as they relate to an IR condition, the mRNA expressions of adipose and muscle RAS components and immuno-relevant molecules were measured along with plasma RAS components. Mean plasma renin activity and Ang II concentrations increased by 89 and 1658%, respectively, while plasma angiotensinogen (AGT) decreased by 49% over the fast, indicative of systemic RAS activation. Prolonged fasting was associated with decreases in adipose and muscle AGT mRNA expressions of 69 and 68%, respectively, corresponding with decreases in tissue protein content, suggesting suppression of local AGT production. Muscle TNF-α mRNA and protein increased by 239 and 314%, whereas those of adipose Acrp30 decreased by 32 and 98%, respectively. Collectively, this study suggests that prolonged fasting activates a systemic RAS, which contributes to an increase in muscle TNF-α and suppression of adipose Acrp30. This targeted and tissue-specific regulation of TNF-α and Acrp30 is likely coordinated to synergistically contribute to the development of an IR-like condition, independent of local RAS activity. These data enhance our understanding of the adaptive mechanisms evolved by elephant seals to tolerate potentially detrimental conditions.

Coping with physiological oxidative stress: a review of antioxidant strategies in seals.

While diving, seals are exposed to apnea-induced hypoxemia and repetitive cycles of ischemia/reperfusion. While on land, seals experience sleep apnea, as well as prolonged periods of food and water deprivation. Prolonged fasting, sleep apnea, hypoxemia and ischemia/reperfusion increase oxidant production and oxidative stress in terrestrial mammals. In seals, however, neither prolonged fasting nor apnea-induced hypoxemia or ischemia/reperfusion increase systemic or local oxidative damage. The strategies seals evolved to cope with increased oxidant production are reviewed in the present manuscript. Among these strategies, high antioxidant capacity and the oxidant-mediated activation of hormetic responses against hypoxia and oxidative stress are discussed. In addition to expanding our knowledge of the evolution of antioxidant defenses and adaptive responses to oxidative stress, understanding the mechanisms that naturally allow mammals to avoid oxidative damage has the potential to advance our knowledge of oxidative stress-induced pathologies and to enhance the translative value of biomedical therapies in the long term.

Effect of induction dose, lactation stage and body condition on tiletamine-zolazepam anaesthesia in adult female grey seals (Halichoerus grypus) under field conditions.

Tiletamine-zolazepam (TZ) was used at a mean (sd) dose of 1.18 (0.15) mg/kg administered intramuscularly to anaesthetise adult female grey seals (Halichoerus grypus) under field conditions at three different stages during their lactation period. A significant correlation was observed between the induction dose and time to induction (r=-0.582, P=0.011). Stage of lactation had a significant effect on condition index (CI), calculated as axial girth divided by length (P<0.001), and time to induction (P=0.009). No effect of CI on induction or recovery time was demonstrated. Respiratory rate decreased during induction and increased significantly (P<0.001) during surgical biopsy of blubber. Recovery occurred after 32.5 (11.9) minutes. Minor complications (tremor, vocalisation and mild dyspnoea) were observed in a small number of cases, none of which required treatment.

Selective brain cooling and its vascular basis in diving seals.

Brain (T(brain)), intra-aorta (T(aorta)), latissimus dorsi muscle (T(m)) and rectal temperature (T(r)) were measured in harp (Pagophilus groenlandicus) and hooded (Cystophora cristata) seals during experimental dives in 4 degrees C water. The median brain cooling was about 1 degrees C during 15 min diving, but in some cases it was as much as 2.5 degrees C. Cooling rates were slow for the first couple of minutes, but increased significantly after about 5 min of diving. The onset of cooling sometimes occurred before the start of the dive, confirming that the cooling is under cortical control, like the rest of the diving responses. T(aorta) also fell significantly, and was always lower than T(brain), while T(m) was fairly stable during dives. Detailed studies of the vascular anatomy of front flippers revealed that brachial arterial blood can be routed either through flipper skin capillaries for nutritive purposes and return through sophisticated vascular heat exchangers to avoid heat loss to the environment, or, alternatively, through numerous arterio-venous shunts in the skin and return by way of large superficial veins, which then carry cold blood to the heart. In the latter situation the extent to which the brain is cooled is determined by the ratio of carotid to brachial arterial blood flow, and water temperature, and the cooling is selective in that only those organs that are circulated will be cooled. It is concluded that T(brain) is actively down-regulated during diving, sometimes by as much as 2.5 degrees C, whereby cerebral oxygen requirements may be reduced by as much as 25% during extended dives.