Postweaning Fast Research Articles

Oxylipin responses to fasting and insulin infusion in a large mammalian model of fasting-induced insulin resistance, the northern elephant seal.

The prolonged, postweaning fast of northern elephant seal (Mirounga angustirostris) pups is characterized by a reliance on lipid metabolism and reversible, fasting-induced insulin resistance, providing a unique model to examine the effects of insulin on lipid metabolism. We have previously shown that acute insulin infusion induced a shift in fatty acid metabolism dependent on fasting duration. This study complements the previous study by examining the effects of fasting duration and insulin infusion on circulating levels of oxylipins, bioactive metabolites derived from the oxygenation of polyunsaturated fatty acids. Northern elephant seal pups were studied at two postweaning periods (n = 5/period): early fasting (1-2 wk postweaning; 127 ± 1 kg) and late fasting (6-7 wk postweaning; 93 ± 4 kg). Different cohorts of pups were weighed, sedated, and infused with 65 mU/kg of insulin. Plasma was collected prior to infusion (T0) and at 10, 30, 60, and 120 min postinfusion. A profile of ∼80 oxylipins was analyzed by UPLC-ESI-MS/MS. Nine oxylipins changed between early and late fasting and eight were altered in response to insulin infusion. Fasting decreased prostaglandin F2α (PGF2α) and increased 14,15-dihydroxyicosatrienoic acid (14,15-DiHETrE), 20-hydroxyeicosatetraenoic acid (20-HETE), and 4-hydroxy-docosahexaenoic acid (4-HDoHE) (P < 0.03) in T0 samples, whereas insulin infusion resulted in an inverse change in area-under-the-curve (AUC) levels in these same metabolites (P < 0.05). In addition, 12-12-hydroperoxyeicosatetraenoic acid (HpETE) and 12-HETE decreased with fasting and insulin infusion, respectively (P < 0.04). The oxylipins altered during fasting and in response to insulin infusion may contribute to the manifestation of insulin resistance and participate in the metabolic regulation of associated cellular processes.

Exogenous GLP-1 stimulates TCA cycle and suppresses gluconeogenesis and ketogenesis in late-fasted northern elephant seals pups

The post-weaning fast of northern elephant seal pups is characterized by a lipid-dependent metabolism and associated with a decrease in plasma glucagon-like peptide-1 (GLP-1), insulin, and glucose and increased gluconeogenesis (GNG) and ketogenesis. We have also demonstrated that exogenous GLP-1 infusion increased plasma insulin despite simultaneous increases in cortisol and glucagon, which collectively present contradictory regulatory stimuli of GNG, ketogenesis, and glycolysis. To assess the effects of GLP-1 on metabolism using primary carbon metabolite profiles in late-fasted seal pups, we dose-dependently infused late-fasted seals with low (LDG; 10 pM/kg; n=3) or high (HDG; 100 pM/kg; n=4) GLP-1 immediately following a glucose bolus (0.5g/kg), using glucose without GLP-1 as control (n=5). Infusions were performed in similarly aged animals 6-8 weeks into their post-weaning fast. The plasma metabolome was measured from samples collected at 5 time points just prior to and during the infusions, and network maps constructed to robustly evaluate the effects of GLP-1 on primary carbon metabolism. HDG increased key tricarboxylic acid (TCA) cycle metabolites, and decreased phosphoenolpyruvate and acetoacetate (P<0.05) suggesting that elevated levels of GLP-1 promote glycolysis and suppress GNG and ketogenesis, which collectively increase glucose clearance. These GLP-1-mediated effects on cellular metabolism help to explain why plasma GLP-1 concentrations decrease naturally in fasting pups as an evolved mechanism to help conserve glucose during the late-fasting period.

In vitro Lipolysis and Leptin Production of Elephant Seal Blubber Using Precision-Cut Adipose Tissue Slices.

Adipose tissue plays key roles in energy homeostasis. Understanding its metabolism and regulation is essential to predict the impact of environmental changes on wildlife health, especially in fasting-adapted species. However, in vivo experimental work in wild vertebrates can be challenging. We have developed a novel in vitro approach of precision-cut adipose tissue slices from northern elephant seal (Mirounga angustirostris) as a complementary approach to whole animal models. Blubber biopsies were collected from 14 pups during early and late post-weaning fast (Año Nuevo, CA, United States), precision-cut into 1 mm thick slices and maintained in culture at 37°C for at least 63 h. The slices exhibited an efficient response to ß-adrenergic stimulation, even after 2 days of culture, revealing good in vitro tissue function. The response to lipolytic stimulus did not vary between regions of outer and inner blubber, but was higher at early than at late fast for inner blubber slices. At early fast, lipolysis significantly reduced leptin production. At this stage, inner blubber slices were also more efficient at producing leptin than outer blubber slices, especially in the non-lipolytic condition. This model will aid the study of adipose tissue metabolism and its response to environmental stressors in marine mammals.

Changes in total mercury, methylmercury, and selenium blood levels during different life history stages of the Baltic grey seal (Halichoerus grypus grypus)

Using the blood of grey seal pups, the blood and milk of female grey seals inhabiting the Hel Marine Station of Gdansk University's Institute of Oceanography (HMS), we monitored the transfer of total mercury (THg), methylmercury (MeHg), and selenium (Se) with blood during foetal life and nursing. Changes in the concentration of mercury and selenium were characterised in the pups' blood during their first three months of life when they transition from suckling, to a post-weaning fast, to eating fish.In the blood of pregnant females, there was a significant decrease in THg and MeHg concentrations throughout the gestation, indicating the transfer of these toxins through the placenta into the foetus. At no other stage of the pup's development was there such a high level of THg and MeHg as on the day of birth, despite the incorporation of mercury into the lanugo during foetal growth. This suggests that the maternal transfer of mercury during gestation may be the time of greatest mercury exposure for a young seal pup. The consumption of milk caused a rapid increase in weight and a lowering of the mercury level in the blood in the subsequent days of the pups' life. The postweaning fast was the period of the lowest mercury concentration. The switch to a diet consisting of fish caused a systematic increase in the concentration of mercury in the blood of the pups.Milk was the significant source of selenium for pups and the selenium concentration in females' blood was reduced during lactation.The nursing period seemed to have the greatest impact on the mercury and selenium blood levels in examined seals. Natural development of the grey seal pup created an opportunity to decrease the levels of toxic substances obtained through the maternal transfer during foetal growth.

Glucocorticoid‐Receptor Mediated Signaling Regulates Lipolysis in Northern Elephant Seal Pups

Northern elephant seals (NES) experience prolonged 2–3 month fasting periods, relying primarily on the oxidation of fatty acids to meet their energetic needs (RQ=0.71). Although fasting periods are characterized by an increase in circulating cortisol (F) and non‐esterified fatty acids (NEFA's), the functional relevance such increase in cortisol in NES is not defined. We hypothesized that F lipolysis is regulated through glucocorticoid receptor signaling (GR) in NES. The contributions of F and the GR were assessed by exogenous infusions of ACTH and/or concurrent blockade of the GR in the following groups: (1) untreated control, (2) ACTH (80 units intramuscular injection (IM)) and (3) ACTH + GR blocker (RU486) (80 units IM + 400mg RU486 in time release pellets). Plasma, and adipose and muscle biopsies were collected at days 0 (T0; immediately prior to infusion) and 6 (T6). Mean plasma NEFA levels increased 38% (0.84 ± 0.13 vs 1.16 ± 0.06mM) from T0 to T6 with ACTH infusion suggesting glucocorticoids contribute to increased lipolysis. ACTH infusion increased mean adipose HSP90 expression by 92%±18.2 while decreasing mean muscle HSP90 expression by 39%±14.5 from T0 to T6 suggesting that HSP90 may participate in GR‐mediated lipolysis in adipose while contributing to conserving lipid sources in muscle. Mean muscle CD36 expression decreased 46%±12.2 from T0 to T6 with ACTH infusion suggesting that there is a reduced intake of NEFA's by skeletal muscle, contributing to increased circulating NEFAs during lipolysis. These data suggest that the fasting‐associated increase in GR signaling is imperative for sustaining elevated levels of lipolysis during the post weaning fast, and is dependent upon the functional role of cortisol and HSP90.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

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.

Making it through the first year: Ontogeny of movement and diving behavior in harbor seals from Svalbard, Norway

AbstractPhocid seal pups must learn successful survival strategies, largely independently, following their abrupt weaning at a relatively young age. To explore the ontogeny of aquatic skills, space use and first‐year habitat choices made by harbor seals, pups (n = 30) were instrumented with satellite relay data loggers (SRDLs) in Svalbard, Norway in 2009 and 2010. Initially, the pups had small home ranges and showed rapid changes in their activity budgets and diving capabilities, displaying steep linear increases in diving depth and duration and in the amount of time spent diving. Most pups underwent an abrupt shift in movement patterns at ca. 50 d of age, which likely marked the end of the postweaning fast. Around this same time, the steep progression in diving performance slowed, though longer, deeper dives gradually became the norm. However, bottom time, ascent and descent rates, and postdive recovery times remained stable after the postweaning fast, suggesting that most aquatic skill acquisition was completed during the first months of life. Few clear effects of environmental variables such as upwelling phenomenon, which are known to influence the diving behavior of adults from the same population, were detected in the diving patterns of pups.

Endocrine response to realimentation in young northern elephant seals (Mirounga angustirostris): Indications for development of fasting adaptation

Most organisms undergo changes in their environment, both predictably and unpredictably, which require them to alter priorities in nutrient allocation with regards to food availability. Species that more predictably encounter extended periods of limited food resources or intake while mitigating the negative effects of starvation are considered to be fasting adapted. Northern elephant seals (NES) are one such species and routinely undergo extended periods of fasting for breeding, molting, as well as a post-weaning fast at 6–8weeks of age. However, during unusual times of nutritional deprivation, animals may enter stage III fasting. While fasting and foraging in this species has been extensively studied, realimentation following fasting beyond normal life history parameters has not been investigated. In this study, changes in ghrelin, growth hormone (GH), and insulin-like growth factor (IGF)-I were compared across 8weeks of realimentation following emaciation in three age classes: neonates, post-molt pups, and yearlings. Longitudinal changes in hormone profiles indicate that neonate and post-molt pups are slow to recover mass and positive energy balance despite an energy dense diet fed at 10% body mass. In addition, ghrelin and GH concentrations remained elevated in post-molt pups compared to other age classes. Changes in hormone concentrations early in realimentation indicate that yearling animals recover more rapidly from periods of nutritional deprivation than do younger animals. Overall, this suggests that the ability to regulate metabolic homeostasis with regards to nutrient allocation may develop over time, even in a species that is considered to be fasting adapted.

Lipophilicity of PCBs and fatty acids determines their mobilisation from blubber of weaned northern elephant seal pups

Polychlorinated biphenyls (PCBs) exhibit lipophilic properties that lead to their bioaccumulation in adipose tissue. Following PCB exposition, northern elephant seals (Mirounga angustirostris) concentrate high amounts of these pollutants in their large adipose tissue stores. During lipolytic periods such as the post-weaning fast, fatty acids (FAs), which form triglycerides, and PCBs are both mobilised from adipose tissue. Our results showed that the degree of lipophilicity of FAs and PCBs impacted their release: the more lipophilic FAs and PCBs tended to be more conserved in blubber over the fast than the less lipophilic ones. This led to an enrichment of more lipophilic compounds within adipocytes with the progression of the fast. Life history patterns that include fasting may thus influence the profile of blubber lipids and contaminants.

Development of Dive Capacity in Northern Elephant Seals (Mirounga angustirostris): Reduced Body Reserves at Weaning Are Associated with Elevated Body Oxygen Stores during the Postweaning Fast.

Developmental increases in dive capacity have been reported in numerous species of air-breathing marine vertebrates. Previous studies in juvenile phocid seals suggest that increases in physiological dive capacity during the postweaning fast (PWF) are critical to support independent aquatic foraging. Although there is a strong relationship between size at weaning and PWF duration and body reserves at weaning vary considerably, few studies have considered whether such variation in body reserve magnitude promotes phenotypic modulation of dive capacity development during the PWF. Phenotypic modulation, a form of developmental plasticity in which rates and degrees of expression of the developmental program are modulated by environmental factors, may enhance diving capacity in weanlings with reduced PWF durations due to smaller body reserves at weaning if reduced body reserves promote accelerated development of dive capacity. We longitudinally measured changes in blood and muscle oxygen stores and muscle metabolic enzymes over the first 8 wk of the PWF in northern elephant seals and determined whether rates of change in these parameters varied with body reserves at weaning. We assessed whether erythropoietin (EPO), thyroid hormones, serum nonesterified fatty acid levels, and iron status influenced blood and muscle oxygen store development or were influenced by body reserves at weaning. Although mass-specific plasma volume and blood volume were relatively stable across the fast, both were elevated in animals with reduced body reserves. Surprisingly, hemoglobin and mean corpuscular hemoglobin concentrations declined over the PWF while hematocrit remained stable, and these variables were not associated with body reserves or EPO. Swimming muscle myoglobin and serum iron levels increased rapidly early in the PWF and were not related to body reserves. Patterns in maximal activities of muscle enzymes suggested a decline in total aerobic and anaerobic metabolic capacity over the PWF, despite maintenance of fat oxidation capacity. These results suggest that only development of blood volume is increased in smaller weanlings and that extended fasting durations in larger weanlings do not improve physiological dive capacity.

Adiponectin and Insulin in Gray Seals during Suckling and Fasting: Relationship with Nutritional State and Body Mass during Nursing in Mothers and Pups.

Animals that fast during breeding and/or development, such as phocids, must regulate energy balance carefully to maximize reproductive fitness and survival probability. Adiponectin, produced by adipose tissue, contributes to metabolic regulation by modulating sensitivity to insulin, increasing fatty acid oxidation by liver and muscle, and promoting adipogenesis and lipid storage in fat tissue. We tested the hypotheses that (1) circulating adiponectin, insulin, or relative adiponectin gene expression is related to nutritional state, body mass, and mass gain in wild gray seal pups; (2) plasma adiponectin or insulin is related to maternal lactation duration, body mass, percentage milk fat, or free fatty acid (FFA) concentration; and (3) plasma adiponectin and insulin are correlated with circulating FFA in females and pups. In pups, plasma adiponectin decreased during suckling (linear mixed-effects model [LME]: T = 4.49; P < 0.001) and the early postweaning fast (LME: T = 3.39; P = 0.004). In contrast, their blubber adiponectin gene expression was higher during the early postweaning fast than early in suckling (LME: T = 2.11; P = 0.046). Insulin levels were significantly higher in early (LME: T = 3.52; P = 0.004) and late (LME: T = 6.99; P < 0.001) suckling than in fasting and, given the effect of nutritional state, were also positively related to body mass (LME: T = 3.58; P = 0.004). Adiponectin and insulin levels did not change during lactation and were unrelated to milk FFA or percentage milk fat in adult females. Our data suggest that adiponectin, in conjunction with insulin, may facilitate fat storage in seals and is likely to be particularly important in the development of blubber reserves in pups.

Mobilisation of blubber fatty acids of northern elephant seal pups (Mirounga angustirostris) during the post-weaning fast

Northern elephant seal pups were longitudinally sampled at Año Nuevo State Reserve during the post-weaning fast, in order to evaluate the changes of fatty acid (FA) profiles in serum as well as in the inner and outer layers of blubber. The major FAs of inner and outer blubber layers were broadly similar to those found in NES maternal milk previously measured, suggesting a direct deposit of dietary FAs in the blubber during the suckling period. The outer blubber layer contained more medium-chain monounsaturated FAs that contribute in keeping the fluidity of this tissue at cold temperatures. It was compensated by higher proportions of saturated FAs in the inner blubber layer. The FA signature of inner blubber, the layer that is mainly mobilised during energy deprivation, slightly differed from the signature of serum. There were greater proportions of medium-chain saturated FAs and ω-6 polyunsaturated FAs, and lower proportions of long-chain saturated FAs, medium-chain monounsaturated FAs and long-chain monounsaturated FAs in serum as compared to inner blubber. We also demonstrated that lipophilicity is the main factor governing the mobilisation of FAs from blubber. The least lipophilic FAs were preferentially hydrolysed from blubber, leading to an enrichment of the more lipophilic FAs in this tissue with the progression of the fast. The expression levels of HSL and ATGL, which are two enzymes involved in the lipolytic process, remained stable during the post-weaning fast. This suggests that the pups have developed the enzymatic mechanisms for an efficient lipolysis as soon as the first week of fast.

New insights into the cardiorespiratory physiology of weaned southern elephant seals (Mirounga leonina).

Southern elephant seal (Mirounga leonina) pups must strike a balance between conserving energy during their post-weaning fast and simultaneously developing diving abilities to attain nutritional independence. Little is known about environmental influences on cardiorespiratory patterns, hence energy use, throughout the 6 week fast. Continuous heart rates were recorded for free-ranging, newly weaned southern elephant seals using heart rate time-depth recorders for 5-9 days at Sub-Antarctic Macquarie Island, during October 1994 (n = 1), 1995 (n = 4) and 1996 (n = 1). Daytime observations of respiration and behaviour were made throughout. We present the first instance of synchronous heart rate traces recorded simultaneously for individual weaners. Generalized additive models revealed that a sinusoidal pattern of diurnal heart rate elevation and nocturnal depression was evident in all seals and, on at least one occasion, a conspicuous break in this pattern coincided with an extreme cold weather event. Seals in this study were capable of considerable cardiorespiratory control and regularly demonstrated bradycardia during periods of resting apnoea. Apnoeic duration ranged from 33 to 291 s (mean 134 s). Apnoeic heart rates (mean 67 ± 15 beats min(-1), range 40-114 beats min(-1)) were on average 19.7% lower than those exhibited during periods of eupnoea (mean 83 ± 15 beats min(-1), range 44-124 beats min(-1)). The early development of the cardiorespiratory response is characterized by arrhythmic heart and respiration rates. The strong temporal patterns observed are being driven by the opposing requirements of maximizing time spent fasting in order to develop diving capabilities and of maximizing departure mass. This pilot study has highlighted a potentially large effect of ambient weather conditions on newly weaned southern elephant seal cardiorespiratory activity. Given the increasing westerlies and more erratic and increasing storminess associated with the Southern Annular Mode predicted in the Southern Ocean, the patterns observed here warrant further investigation.

Mobilisation of lipophilic pollutants from blubber in northern elephant seal pups (Mirounga angustirostris) during the post-weaning fast

Northern elephant seals (NES) (Mirounga angustirostris) from the Año Nuevo State Reserve (CA, USA) were longitudinally sampled during the post-weaning fast in order to study the mobilisation and redistribution of various classes of persistent organic pollutants (POPs), such as polybrominated diphenyl ethers (PBDEs), polychlorinated biphenyls (PCBs), dichlorodiphenyldichloroethylene (p,p′-DDE) and hexachlorobenzene (HCB) between blubber and blood. Inner and outer blubber layers were analysed separately. Organohalogenated compounds were detected in all blubber samples in the decreasing order of their concentrations: p,p′-DDE>PCBs⪢HCB>PBDEs. The concentrations of all studied compounds were homogeneously distributed in the blubber layer at early fast, since the concentrations of POPs were statistically not different in the inner and outer layers. With the progression of the fast, the concentrations of PBDEs, PCBs and p,p′-DDE increased more sharply in inner blubber than in outer blubber. As a result, their levels became significantly higher in inner blubber as compared to outer blubber at late fast. The rise of pollutant concentrations in blubber might result from a less efficient mobilisation than triglycerides and/or a reuptake by adipocytes of some of the pollutants released into the circulation. The mobilisation of pollutants from blubber was higher at late fast. An increase of pollutant concentrations was observed in serum between early and late fast. Lower halogenated congeners (i.e. tetra-CBs) were present in higher proportions in serum, whereas the higher halogenated congeners (i.e. hepta-CBs) were mainly found in the inner and outer blubber layers. The transfer ratios of both PBDEs and PCBs from inner blubber to serum decreased with the number of chlorine and bromine atoms. In addition, the distribution of both types of compounds between serum and blubber was strongly influenced by their lipophilic character (logKow values), with more lipophilic compounds being less efficiently released from blubber to serum.

The development of dive capacity in northern elephant seal pups during the post‐weaning fast (1100.14)

The post‐weaning fast (PWF) in seals has been associated with development of the physiological and behavioral capacity to perform the breath‐hold dives which characterize at‐sea behavior in these species. Previous studies in juvenile northern elephant seals (Mirounga angustirostris) indicate that the duration of the PWF varies substantially with body reserves at weaning, and many suggest that the terrestrial PWF enables juveniles to develop the physiological capacity to dive before initiating independent foraging at sea. However, few studies have examined the effect of magnitude of body reserves at weaning on development of dive capacity. We measured energy reserves at weaning and made longitudinal measurements of components of blood and muscle oxygen stores, thyroid and glucocorticoid hormones, iron status markers, muscle metabolic enzyme activities, and field metabolic rate in northern elephant seal pups during the first 8 weeks of the PWF. Our objective was to assess the impacts of body reserves at weaning on the rate of development during the first 8 weeks of the PWF. Although myoglobin concentration, a critical determinant of the muscle oxygen store, increases during the first eight weeks of the PWF, other key components of aerobic diving capacity do not increase during this same time period. When the magnitude of body reserves at weaning impacted the measured parameters of aerobic dive capacity, pups with smaller body reserves at weaning tended to exhibit greater cumulative increases across the PWF. Pups with smaller body reserves at weaning develop greater mass specific oxygen storage capacity during the first 8 weeks of the PWF. However, the greatest changes in dive capacity may occur near the end of the fast or after juveniles have left the natal rookery to initiate independent foraging.Grant Funding Source: Supported by NSF

Characterization of blubber fatty acid signatures in northern elephant seals (Mirounga angustirostris) over the postweaning fast

Phocids routinely fast for extended periods. During these fasts, energetic requirements are met primarily through the catabolism of blubber lipid. To assess whether fatty acid (FA) composition changes during the postweaning fast in northern elephant seals, blubber biopsies were acquired longitudinally from 43 pups at 2.3±1.5 and 55.2±3.7days postweaning in 1999 and 2000. At weaning, short-chain monounsaturated FA (SC-MUFA, ≤18 carbons) dominated the blubber while saturated FA (SFA) were found in the next highest proportion. The major FA (all ≥1% by mass) comprised approximately 91% of total blubber FA. In both years, 18:1n-9 and 16:0 were the most prevalent FA. Major FA mobilized during the fast consisted of polyunsaturated FA (PUFA), SFA, and SC-MUFA. Long-chain MUFA (>18 carbons) tended to be conserved. The fractional mobilization value of 20:5n-3 was the highest, resulting in significant reductions of this PUFA. Although concentrations of some blubber FA changed significantly during the postweaning fast, the general FA signature of blubber was similar at weaning and near the end of the fast. Changes in some FA differed across years. For example, the concentration of 20:4n-6, a minor PUFA, was significantly reduced in 1999 but not in 2000. FA mobilization patterns in northern elephant seal pups are somewhat similar to those reported previously for other fasting phocids and terrestrial mammals, though there are some notable differences. Differences in FA mobilization patterns across mammalian species may be related to differences in diets, geographical distribution, environmental factors, physiological adaptations, and life history stage.

A profile of carbohydrate metabolites in the fasting northern elephant seal

Northern elephant seals endure prolonged periods of food deprivation at multiple life-history stages and simultaneous with energetically costly activities—including reproduction and development. Most mammals decrease their energy expenditure while fasting, with simultaneous reductions in gluconeogenesis and circulating glucose concentration. Paradoxically, elephant seals maintain high rates of both energy expenditure and gluconeogenesis, and high blood glucose concentrations throughout fasting. We therefore characterized the suite of changes that occur in carbohydrate metabolites during fasting in northern elephant seals. Using a broad-based metabolomics platform we investigated fasting during two states—lactation in adult females and the post-weaning developmental period in pups. A total of 227 metabolites were detected in seal plasma; 31 associated with carbohydrate metabolism were analyzed in the present study. Several compounds showed similar responses during lactation and the post-weaning fast (e.g. glycerol and mesaconate) whereas other compounds displayed quite different abundances between groups (e.g. citrate and pyruvate). This work found that, while the changes that occur with fasting were frequently similar in lactating females and developing pups, the relative abundance of compounds often varied markedly. These differences suggest that the metabolic strategies used to endure prolonged fasts are influenced by life-history or nutrient constraints.

Activation of systemic, but not local, renin-angiotensin system is associated with up-regulation 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.

How are trace elements mobilized during the postweaning fast in Northern elephant seals?

Northern elephant seal (Mirounga angustirostris) pups undergo a substantial intertissue reorganization of protein, minerals, and other cellular components during their postweaning development, which might entail the mobilization of associated contaminants. The authors investigated the changes in concentrations of 11 elements (Ca, Cd, Cr, Cu, Fe, Hg, Ni, Pb, Se, V, and Zn) in a longitudinal study on 22 northern elephant seal pups during the postweaning fast. Slight changes in most element concentrations were observed in blood throughout the fast. Circulating levels of Hg, Se, and Cu appeared less altered during the postweaning fast than measured during suckling. Despite the considerable fat utilization, element concentrations, except Fe, in blubber remained stable throughout the fast, which suggests that elements are mobilized from blubber as efficiently as lipids. As indicators of the placental transfer, concentrations in lanugo hair revealed the existence of maternal transfer and accumulation of all assayed trace elements during fetal development. In addition, the new pelage, rapidly produced after weaning, appeared to be an important elimination route for toxic metals such as Hg, Cd, and Pb. The high mineral content detected in pup hair suggests that this species would be more exposed to trace elements than other phocids (except Cd and Pb). Nevertheless, this statement needs further monitoring and toxicological studies to determine better the exposition to trace elements and its potential impact on the health of the northern elephant seal.

Decreased expression of adipose CD36 and FATP1 are associated with increased plasma non-esterified fatty acids during prolonged fasting in northern elephant seal pups (Mirounga angustirostris)

The northern elephant seal pup (Mirounga angustirostris) undergoes a 2-3 month post-weaning fast, during which it depends primarily on the oxidation of fatty acids to meet its energetic demands. The concentration of non-esterified fatty acids (NEFAs) increases and is associated with the development of insulin resistance in late-fasted pups. Furthermore, plasma NEFA concentrations respond differentially to an intravenous glucose tolerance test (ivGTT) depending on fasting duration, suggesting that the effects of glucose on lipid metabolism are altered. However, elucidation of the lipolytic mechanisms including lipase activity during prolonged fasting in mammals is scarce. To assess the impact of fasting and glucose on the regulation of lipid metabolism, adipose tissue and plasma samples were collected before and after ivGTTs performed on early (2 weeks, N=5) and late (6-8 weeks; N=8) fasted pups. Glucose administration increased plasma triglycerides and NEFA concentrations in late-fasted seals, but not plasma glycerol. Fasting decreased basal adipose lipase activity by 50%. Fasting also increased plasma lipase activity twofold and decreased the expressions of CD36, FAS, FATP1 and PEPCK-C by 22-43% in adipose tissue. Plasma acylcarnitine profiling indicated that late-fasted seals display higher incomplete LCFA β-oxidation. Results suggest that long-term fasting induces shifts in the regulation of lipolysis and lipid metabolism associated with the onset of insulin resistance in northern elephant seal pups. Delineation of the mechanisms responsible for this shift in regulation during fasting can contribute to a more thorough understanding of the changes in lipid metabolism associated with dyslipidemia and insulin resistance in mammals.