Diversified regulation of circadian clock gene expression following whole genome duplication.

Across taxa, circadian control of physiology and behavior arises from cell-autonomous oscillations in gene expression, governed by a networks of so-called 'clock genes', collectively forming transcription-translation feedback loops. In modern vertebrates, these networks contain multiple copies of clock gene family members, which arose through whole genome duplication (WGD) events during evolutionary history. It remains unclear to what extent multiple copies of clock gene family members are functionally redundant or have allowed for functional diversification. We addressed this problem through an analysis of clock gene expression in the Atlantic salmon, a representative of the salmonids, a group which has undergone at least 4 rounds of WGD since the base of the vertebrate lineage, giving an unusually large complement of clock genes. By comparing expression patterns across multiple tissues, and during development, we present evidence for gene- and tissue-specific divergence in expression patterns, consistent with functional diversification of clock gene duplicates. In contrast to mammals, we found no evidence for coupling between cortisol and circadian gene expression, but cortisol mediated non-circadian regulated expression of a subset of clock genes in the salmon gill was evident. This regulation is linked to changes in gill function necessary for the transition from fresh- to sea-water in anadromous fish. Overall, this analysis emphasises the potential for a richly diversified clock gene network to serve a mixture of circadian and non-circadian functions in vertebrate groups with complex genomes.

Arctic charr brain transcriptome strongly affected by summer seasonal growth but only subtly by feed deprivation.

BACKGROUND: The Arctic charr (Salvelinus alpinus) has a highly seasonal feeding cycle that comprises long periods of voluntary fasting and a short but intense feeding period during summer. Therefore, the charr represents an interesting species for studying appetite-regulating mechanisms in fish. RESULTS: In this study, we compared the brain transcriptomes of fed and feed deprived charr over a 4 weeks trial during their summer feeding season. Despite prominent differences in body condition between fed and feed deprived charr at the end of the trial, feed deprivation affected the brain transcriptome only slightly. In contrast, the transcriptome differed markedly over time in both fed and feed deprived charr, indicating strong shifts in basic cell metabolic processes possibly due to season, growth, temperature, or combinations thereof. The GO enrichment analysis revealed that many biological processes appeared to change in the same direction in both fed and feed deprived fish. In the feed deprived charr processes linked to oxygen transport and apoptosis were down- and up-regulated, respectively. Known genes encoding for appetite regulators did not respond to feed deprivation. Gene expression of Deiodinase 2 (DIO2), an enzyme implicated in the regulation of seasonal processes in mammals, was lower in response to season and feed deprivation. We further found a higher expression of VGF (non-acronymic) in the feed deprived than in the fed fish. This gene encodes for a neuropeptide associated with the control of energy metabolism in mammals, and has not been studied in relation to regulation of appetite and energy homeostasis in fish. CONCLUSIONS: In the Arctic charr, external and endogenous seasonal factors for example the increase in temperature and their circannual growth cycle, respectively, evoke much stronger responses in the brain than 4 weeks feed deprivation. The absence of a central hunger response in feed deprived charr give support for a strong resilience to the lack of food in this high Arctic species. DIO2 and VGF may play a role in the regulation of energy homeostasis and need to be further studied in seasonal fish.

Lifelong Exposure to PCBs in the Remote Norwegian Arctic Disrupts the Plasma Stress Metabolome in Arctic Charr.

Lake Ellasjøen on the remote Norwegian island of Bjørnøya is populated by Arctic charr (Salvelinus alpinus) having 20-fold higher body burdens of polychlorinated biphenyls (PCB) compared to charr from the neighboring Lake Laksvatn. This provides a natural setting to test the hypothesis that lifelong exposure to PCBs compromises the energy metabolism in this northernmost living salmonid. To test this, blood was sampled from charr from both lakes immediately after capture and following a 1 h handling and confinement stressor to assess possible differences in their energy metabolism and energy substrate mobilization, respectively. The plasma metabolome of charr was assessed by metabolite detection/separation with LC-MS. Plasma metabolite profiles revealed differences in key pathways involved in amino acid metabolism between charr from each lake, underscoring an impact of PCBs on energy metabolism in Arctic charr residing in Lake Ellasjøen. Subjecting charr from either lake to an acute stressor altered the plasma metabolite profiles and revealed distinct stress metabolome in Lake Ellasjøen charr, suggesting a reduced metabolic capacity. Taken together, lifelong exposure to PCBs in Ellasjøen charr disrupts the plasma metabolome, and may impair the adaptive metabolic response to stressors, leading to a reduced fitness.

Photoperiod revisited: is there a critical day length for triggering a complete parr-smolt transformation in Atlantic salmon Salmo salar?

The present study investigated whether there is a critical length of photoperiod needed to stimulate a completed parr-smolt transformation (PST) in Atlantic salmon Salmo salar. In two experiments, S. salar parr of the Norwegian aquaculture strain held on continuous light were exposed to a short photoperiod (6 L:18D) followed by exposure to 8 L:16D, 12 L:12D, 16 L:8D, 20 L:4D and 24 L:0D in experiment 1 or to 6 L:18D followed by maintenance on 6 L:18D or exposure to 12 L:12D and 24 L:0D photoperiods in experiment 2. All groups, irrespective of photoperiod treatment, developed improved hypo-osmoregulatory ability. However, the development was greatest in the groups exposed to 20 L:4D and 24 L:0D in experiment 1 and 24 L:0D in experiment 2. In experiment 2, gill Na+ - K+ -ATPase activity increased in the group exposed to 24 L:0D, but not in the groups exposed to 12 L:12D and 6 L:18D. The groups exposed to 20 L:4D and 24 L:0D in experiment 1 and 24 L:0D in experiment 2 also grew better than fish exposed to shorter photoperiods. In experiment 2 only the group exposed to 24 L:0D showed a decrease in condition factor and increases in plasma growth hormone and brain type 2 deiodinase mRNA abundance. Hence, only the groups exposed to photoperiods above 16 L:8D developed classical smolt indices in the present experiment, leading us to conclude that the photoperiod increase needs to exceed 16 h daylight for stimulating a complete PST in the S. salar used in the present study.

Exposure to Persistent Organic Pollutants Reduces Testosterone Concentrations and Affects Sperm Viability and Morphology during the Mating Peak Period in a Controlled Experiment on Farmed Arctic Foxes (Vulpes lagopus).

We investigated testosterone production and semen parameters in farmed Arctic foxes by dietary exposure to persistent organic pollutants (POPs) for 22 months. Eight male foxes were given a diet of POP-contaminated minke whale blubber, whereas their eight male siblings were fed a control diet containing pig fat as the main fat source. The minke whale-based feed contained a ∑POPs concentration of 802 ng/g ww, whereas the pig-based feed contained ∑POPs of 24 ng/g ww. At the end of the experiment, ∑POP concentrations in adipose tissue were 8856 ± 2535 ng/g ww in the exposed foxes and 1264 ± 539 ng/g ww in the control foxes. The exposed group had 45-64% significantly lower testosterone concentrations during their peak mating season compared to the controls (p ≤ 0.05), while the number of dead and defect sperm cells was 27% (p = 0.07) and 15% (p = 0.33) higher in the exposed group. Similar effects during the mating season in wild Arctic foxes may affect mating behavior and reproductive success. On the basis of these results, we recommend testosterone as a sensitive biomarker of POP exposure and that seasonal patterns are investigated when interpreting putative endocrine disruption in Arctic wildlife with potential population-level effects.

Feedback from Arctic charr: Feed flavour stimulation and re-feeding after feed deprivation stimulate genes encoding both orexigenic and anorexigenic neuropeptides.

Despite vast research attention, the knowledge about central mechanisms of appetite regulation in teleost remains inconclusive. A common strategy in studies on appetite regulating mechanisms is to measure the response to feed restriction or - deprivation, but responses vary between fish species and between experiments, and are also likely dependent on the degree of energy perturbation. The anadromous Arctic charr is an interesting model for studying appetite regulation as its feeding cycle comprises months of winter anorexia, and hyperphagia during summer. Here we studied how the gene expression of putative hypothalamic appetite regulators were affected by two days, one week and one month feed deprivation during summer, and subsequent re-feeding and exposure to feed flavour. Short-term feed deprivation caused only a minor reduction in condition factor and had no effect on hypothalamic gene expression. Long-term feed-deprivation caused a marked reduction in weight and condition factor which contrasted the increase in weight and condition factor seen in ad libitum fed controls. A marked energy perturbation by feed deprivation was also indicated by a lower hypothalamic expression of the genes encoding insulin-like growth factor 1 (IGF1) and IGF1 binding protein 5 in the feed deprived charr compared to fed controls. Surprisingly, long-term feed deprivation and energy perturbation did not induce changes in hypothalamic appetite regulators. Unexpectedly, re-feeding and exposure to feed flavour caused an increase in the expression of the genes encoding the orexigenic agouti-related peptide and the anorexigenic melanocortin receptor 4 and cocaine- and amphetamine-regulated transcript. Our study gives strong evidence for a role of these in appetite regulation in Arctic charr, but their mechanisms of action remain unknown. We suggest that changes in gene expression are more likely to be registered during transition phases, e.g. from fasting to feeding and upon stimulatory inputs such as feed flavour.

Drastic neofunctionalization associated with evolution of the timezyme AANAT 500 Mya.

Melatonin (N-acetyl-5-methoxytrypamine) is the vertebrate hormone of the night: circulating levels at night are markedly higher than day levels. This increase is driven by precisely regulated increases in acetylation of serotonin in the pineal gland by arylalkylamine N-acetyltransferase (AANAT), the penultimate enzyme in the synthesis of melatonin. This unique essential role of AANAT in vertebrate timekeeping is recognized by the moniker the timezyme. AANAT is also found in the retina, where melatonin is thought to play a paracrine role. Here, we focused on the evolution of AANAT in early vertebrates. AANATs from Agnathans (lamprey) and Chondrichthyes (catshark and elephant shark) were cloned, and it was found that pineal glands and retinas from these groups express a form of AANAT that is compositionally, biochemically, and kinetically similar to AANATs found in bony vertebrates (VT-AANAT). Examination of the available genomes indicates that VT-AANAT is absent from other forms of life, including the Cephalochordate amphioxus. Phylogenetic analysis and evolutionary rate estimation indicate that VT-AANAT evolved from the nonvertebrate form of AANAT after the Cephalochordate-Vertebrate split over one-half billion years ago. The emergence of VT-AANAT apparently involved a dramatic acceleration of evolution that accompanied neofunctionalization after a duplication of the nonvertebrate AANAT gene. This scenario is consistent with the hypotheses that the advent of VT-AANAT contributed to the evolution of the pineal gland and lateral eyes from a common ancestral photodetector and that it was not a posthoc recruitment.

Dietary contaminant exposure affects plasma testosterone, but not thyroid hormones, vitamin A, and vitamin E, in male juvenile arctic foxes (Vulpes lagopus).

Levels of persistent organic pollutants (POP), such as polychlorinated biphenyls (PCB), are high in many Arctic top predators, including the Arctic fox (Vulpes lagopus). The aim of this study was to examine possible endocrine-disruptive effects of dietary POP exposure in male juvenile Arctic foxes in a controlled exposure experiment. The study was conducted using domesticated farmed blue foxes (Vulpes lagopus) as a model species. Two groups of newly weaned male foxes received a diet supplemented with either minke whale (Baleneoptera acutorostrata) blubber that was naturally contaminated with POP (exposed group, n = 5 or 21), or pork (Sus scrofa) fat (control group, n = 5 or 21). When the foxes were 6 mo old and had received the 2 diets for approximately 4 mo (147 d), effects of the dietary exposure to POP on plasma concentrations of testosterone (T), thyroid hormones (TH), thyroid-stimulating hormone (TSH), retinol (vitamin A), and tocopherol (viramin E) were examined. At sampling, the total body concentrations of 104 PCB congeners were 0.1 ± 0.03 µg/g lipid weight (l.w.; n = 5 [mean ± standard deviation]) and 1.5 ± 0.17 µg/g l.w. (n = 5) in the control and exposed groups, respectively. Plasma testosterone concentrations in the exposed male foxes were significantly lower than in the control males, being approximately 25% of that in the exposed foxes. There were no between-treatment differences for TH, TSH, retinol, or tocopherol. The results suggest that the high POP levels experienced by costal populations of Arctic foxes, such as in Svalbard and Iceland, may result in delayed masculine maturation during adolescence. Sex hormone disruption during puberty may thus have lifetime consequences on all aspects of reproductive function in adult male foxes.

Photoperiod-dependent developmental reprogramming of the transcriptional response to seawater entry in Atlantic salmon (Salmo salar).

The developmental transition of juvenile salmon from a freshwater resident morph (parr) to a seawater (SW) migratory morph (smolt), known as smoltification, entails a reorganization of gill function to cope with the altered water environment. Recently, we used RNAseq to characterize the breadth of transcriptional change which takes place in the gill in the FW phase of smoltification. This highlighted the importance of extended exposure to short, winter-like photoperiods (SP) followed by a subsequent increase in photoperiod for completion of transcriptional reprogramming in FW and efficient growth following transfer to SW. Here, we extend this analysis to examine the consequences of this photoperiodic history-dependent reprogramming for subsequent gill responses upon exposure to SW. We use RNAseq to analyze gill samples taken from fish raised on the photoperiod regimes we used previously and then challenged by SW exposure for 24 hours. While fish held on constant light (LL) throughout were able to hypo-osmoregulate during a 24 hours SW challenge, the associated gill transcriptional response was highly distinctive from that in fish which had experienced a 7-week period of exposure to SP followed by a return to LL (SPLL) and had consequently acquired the characteristics of fully developed smolts. Fish transferred from LL to SP, and then held on SP for the remainder of the study was unable to hypo-osmoregulate, and the associated gill transcriptional response to SW exposure featured many transcripts apparently regulated by the glucocorticoid stress axis and by the osmo-sensing transcription factor NFAT5. The importance of these pathways for the gill transcriptional response to SW exposure appears to diminish as a consequence of photoperiod mediated induction of the smolt phenotype, presumably reflecting preparatory developmental changes taking place during this process.

Immunologic Profiling of the Atlantic Salmon Gill by Single Nuclei Transcriptomics.

Anadromous salmonids begin life adapted to the freshwater environments of their natal streams before a developmental transition, known as smoltification, transforms them into marine-adapted fish. In the wild, smoltification is a photoperiod-regulated process, involving radical remodeling of gill function to cope with the profound osmotic and immunological challenges of seawater (SW) migration. While prior work has highlighted the role of specialized "mitochondrion-rich" cells (MRCs) and accessory cells (ACs) in delivering this phenotype, recent RNA profiling experiments suggest that remodeling is far more extensive than previously appreciated. Here, we use single-nuclei RNAseq to characterize the extent of cytological changes in the gill of Atlantic salmon during smoltification and SW transfer. We identify 20 distinct cell clusters, including known, but also novel gill cell types. These data allow us to isolate cluster-specific, smoltification-associated changes in gene expression and to describe how the cellular make-up of the gill changes through smoltification. As expected, we noted an increase in the proportion of seawater mitochondrion-rich cells, however, we also identify previously unknown reduction of several immune-related cell types. Overall, our results provide fresh detail of the cellular complexity in the gill and suggest that smoltification triggers unexpected immune reprogramming.

Chronic dietary exposure to environmental organochlorine contaminants induces thyroid gland lesions in Arctic foxes (Vulpes lagopus).

The impact of dietary organochlorine (OC) exposure on thyroid gland pathology was studied in farmed male Arctic foxes (Vulpes lagopus). The exposed group (n=16) was fed a diet based on wild minke whale (Balaenoptera acutorostrata) blubber as a main fat source in order to mimic the exposure to OC cocktails in the Artic environment. This resulted in an exposure of approximately 17 microg Sigma OC/kg day and a Sigma OC residue adipose tissue and liver concentration of 1700 and 4470 ng/gl.w., respectively, after 16 months of exposure. Control foxes (n=13) were fed a diet with pork (Sus scrofa) fat as a main fat source containing significantly lower OC concentrations. The food composition fed to the control and exposed group was standardized for nutrient contents. Four OC-related histopathological changes were found: (1) flat-epithelial-cell true thyroid cysts (TC) characterized by neutral content; (2) remnants of simple squamous epithelial-cell embryonic ducts containing neutral debris (EDN); (3) remnants of stratified squamous epithelial-cell embryonic ducts containing acid mucins often accompanied with debris of leukocyte inflammatory nature (EDM) and (4) disseminated thyroid C-cell hyperplasia (HPC). Of these, the prevalence of TC, EDN and HPC was significantly highest in the exposed group (chi(2) test: all p<0.04). The study shows that the OC mixture in minke whale blubber may cause development of thyroid gland cysts, C-cell hyperplasia and increase the prevalence of cystic remnants of embryonic ducts. The mechanism causing these effects could include endocrine disruption of the hypothalamus-pituitary-thyroid (HPT) axis, a disturbance of the calcium homeostasis/metabolism or energy metabolism or immune suppression. Because concentrations of OCs are higher in wild Arctic foxes, it is likely that these animals could suffer from similar OC-induced thyroid gland pathological and functional changes.

Growth hormone and cortisol treatment stimulate seawater tolerance in both anadromous and landlocked Arctic charr.

In a comparative experiment the effect of cortisol and growth hormone (GH) on the hypo-osmoregulatory ability of a landlocked and an anadromous strain of Arctic charr (Salvelinus alpinus) was investigated. Cortisol and GH were implanted either alone or in combination, and the fish were exposed to a 24 h seawater challenge test (SWT) on days 14 and 28 after implantation. Hypo-osmoregulatory ability, measured as plasma osmolality and chloride concentration after the SWTs, was better in the anadromous than in the landlocked strain, irrespective of treatment. However, cortisol provided a strong stimulation of hypo-osmoregualtory ability in both strains, and this stimulation seemed to be potentiated by GH in an additive manner. Improved hypo-osmoregulatory ability in GH+cortisol treated anadromous Arctic charr was accompanied by increased gill Na(+), K(+)-ATPase activity and Na(+)-K(+)-2Cl(-) cotransporter protein abundance, but no changes in gill Na(+),K(+)-ATPase alpha1a and alpha1b mRNA levels. For landlocked charr the improved hypo-osmoregulatory ability in GH+cortisol treated fish was accompanied only with an increase in gill Na(+)-K(+)-2Cl(-) cotransporter protein abundance. Hormone treatment caused an improvement of hypo-osmoregulatory ability that was of approximately the same magnitude in the landlocked as in the anadromous Arctic charr. This suggests that the lack of spontaneous development of hypo-osmoregulatory ability often seen in landlocked populations of Arctic charr may depend, at least partly, on a lack of the hormonal activation seen in anadromous populations.

Mandibular shape in farmed Arctic foxes (Vulpes lagopus) exposed to persistent organic pollutants.

We investigated if dietary exposure to persistent organic pollutants (POPs) affect mandibular asymmetry and periodontal disease in paired male-siblings of Arctic foxes (Vulpes lagopus). During ontogeny, one group of siblings was exposed to the complexed POP mixture in naturally contaminated minke whale (Balaenoptere acutorostarta) blubber (n = 10), while another group was given wet feed based on pig (Sus scrofa) fat as a control (n = 11). The ∑POP concentrations were 802 ng/g ww in the whale-based feed compared to 24 ng/g ww in the control diet. We conducted a two-dimensional geometric morphometric (GM) analysis of mandibular shape and asymmetry in the foxes and compared the two groups. The analyses showed that directional asymmetry was higher than fluctuating asymmetry in both groups and that mandibular shape differed significantly between the exposed and control group based on discriminant function analysis (T2 = 58.52, p = 0.04, 1000 permutations). We also found a non-significantly higher incidence of periodontal disease (two-way ANOVA: p = 0.43) and greater severity of sub-canine alveolar bone deterioration similar to periodontitis (two-way ANOVA: p = 0.3) in the POP-exposed group. Based on these results, it is possible that dietary exposure to a complexed POP mixture lead to changes in jaw morphology in Arctic foxes. This study suggests that extrinsic factors, such as dietary exposure to POPs, may affect mandibular shape and health in a way that could be harmful to wild Arctic populations. Therefore, further studies using GM analysis as an alternative to traditional morphometric methods should be conducted for wild Arctic fox populations exposed to environmental contaminants.

Monitoring contaminants from oil production at sea by measuring gill EROD activity in Atlantic cod (Gadus morhua).

An ex vivo gill EROD assay was applied in Atlantic cod (Gadus morhua) as a biomarker for waterborne CYP1A-inducing compounds derived from oil production at sea. Exposure to nominal concentrations of 1 ppm or 10 ppm North Sea crude oil in a static water system for 24 h caused a concentration-dependent gill EROD induction. Further, exposure of cod for 14 days to environmentally relevant concentrations of produced water (PW, diluted 1:200 or 1:1000) from a platform in the North Sea using a flow-through system resulted in a concentration-dependent induction of gill EROD. Crude oil (0.2 ppm) from the same oil field also proved to induce EROD. Finally, gill EROD activity in cod caged for 6 weeks at 500-10 000 m from two platforms outside Norway was measured. The activities in these fish were very low and did not differ from those in fish caged at reference sites.

Elevated spring temperature stimulates growth, but not smolt development, in anadromous Arctic charr.

Parr-smolt transformation and growth were studied in captive offspring of anadromous Arctic charr (Salvelinus alpinus) from the Hals watercourse in northern Norway (70 degrees N), held either at a natural temperature (<1 degrees C until May) or at a temperature elevated to 6 degrees C in late March. In mid-May, 5 weeks after the increase in photoperiod from 8:16 h light:dark to continuous light, gill Na+, K(+)-ATPase activity started to increase in both temperature groups, concurrent with the final development of full seawater tolerance. Temperature had no effect on the development of gill Na+, K(+)-ATPase activity, or on hypoosmoregulatory ability. The fish in both treatments resumed growth in mid-May, but from then on growth was faster in the elevated than in the ambient temperature group. In the former group, fish mass doubled in 6 weeks (from 65 to 137 g), and growth ceased at the time when the fish were about to complete their parr-smolt transformation. These findings show that an early vernal temperature increase advances the seasonal growth cycle, but not the parr-smolt transformation, in anadromous Arctic charr.

Extended fasting does not affect the liver innate immune response in rainbow trout.

Activation of immune response pathway is energy demanding. We tested the hypothesis that negative energy balance will curtail the liver's capacity to evoke an immune response in rainbow trout (Oncorhynchus mykiss). Fish were either fed or fasted for 118 d and challenged with lipopolysaccharide (LPS) to determine the liver capacity to elicit an immune response. Fasting led to negative specific growth rate, reduced tissue metabolite levels, and higher transcript abundance of SOCS-2. LPS treatment increased the liver transcript abundances of IL-1ß and IL-8 and serum amyloid protein A, while SOCS-2 was reduced. LPS lowered plasma cortisol level only in the fasted fish, but did not affect liver glucocorticoid or mineralocorticoid receptor protein expressions. Extended fasting did not suppress the liver capacity to evoke an immune response. Upregulation of liver SOCS-2 may be playing a key role in the energy repartitioning, thereby facilitating immune response activation despite extended fasting in trout.

Halogenated organic contaminants and mercury in northern fulmars (Fulmarus glacialis): levels, relationships to dietary descriptors and blood to liver comparison.

The northern fulmar (Fulmarus glacialis) is an interesting candidate for the study of patterns and levels of halogenated organic contaminants (HOCs) since they accumulate high levels of certain HOCs. In the present study we characterized a suite of established and novel HOCs in northern fulmars breeding on Bjørnøya in the Norwegian Arctic. A comparison between blood and liver HOC levels was made, and the levels were related to the ratios of heavier to lighter stable isotopes of nitrogen (15)N/(14)N (delta(15)N) and carbon (13)C/(12)C (delta(13)C) in muscle. A significant difference in congener patterns between blood and liver was found. The delta(13)C was not related to HOCs, neither in liver nor in blood. Weak correlations were found between delta(15)N and liver HOC levels. The 2,3,7,8-tetrachlorodibenzo-p-dioxin toxic equivalents (TEQs) in northern fulmars were well above thresholds for reproductive effects in seabirds.

NFAT5 genes are part of the osmotic regulatory system in Atlantic salmon (Salmo salar).

The anadromous Atlantic salmon utilizes both fresh and salt water (FW and SW) habitats during its life cycle. The parr-smolt transformation (PST) is an important developmental transition from a FW adapted juvenile parr to a SW adapted smolt. Physiological changes in osmoregulatory tissues, particularly the gill, are key in maintaining effective ion regulation during PST. Changes are initiated prior to SW exposure (preparative phase), and are completed when smolts enter the sea (activational phase) where osmotic stress may directly stimulate changes in gene expression. In this paper we identify 4 nuclear factor of activated T cells (NFAT5, an osmotic stress transcription factor) paralogues in Atlantic salmon, which showed strong homology in characterized functional domains with those identified in other vertebrates. Two of the identified paralogues (NFAT5b1 and NFAT5b2) showed increased expression following transfer from FW to SW. This effect was largest in parr that were maintained under short day photoperiod, and showed the highest increases in chloride ion levels in response to SW exposure. The results of this study suggest that NFAT5 is involved in the osmotic stress response of Atlantic salmon.

Biomarker response and hypothalamus-pituitary-interrenal axis functioning in Arctic charr from Bjørnøya (74°30' N), Norway, with high levels of organohalogenated compounds.

The populations of Arctic charr (Salvelinus alpinus) residing in Lake Ellasjøen at Bjørnøya Island in the Norwegian Arctic (74° 30'N, 19° 00'E) possess substantially higher levels of organohalogenated compounds (strongly dominated by polychlorinated biphenyls, PCBs) than conspecifics residing in other, proximate lakes on the island. In the present study we sampled large (<400g), immature charr from Lake Ellasjøen (high PCB levels) and Lake Laksvatn (reference lake, low PCB levels) by hook and line for an immediate blood sampling, and blood and tissue sampling after a 1h confinement stressor. This was done in order to investigate possible effects of pollutants on an acute stress performance in a high-latitude fish species by comparing muscle PCB levels, hepatic cytochrome P4501A (CYP1A) biomarker activation and functioning of the hypothalamus-pituitary-interrenal (HPI) axis between these two populations of Arctic charr. As expected sum PCB muscle levels were 8-fold higher on a wet weigh basis, and 19-fold higher on a lipid weight basis, in charr from Ellasjøen than in charr from Laksvatn. This was accompanied by a 3.5-fold higher liver cyp1a mRNA abundance in the Ellasjøen charr compared to Laksvatn charr. Brain transcript levels encoding glucocorticoid receptor 1 and 2 (GR2) and corticotropin-releasing factor, and pituitary transcript levels encoding GR2 and proopiomelanocortin A1 and A2 were higher in Ellasjøen charr than in Laksvatn charr, while interrenal transcript levels encoding melanocortin 2 receptor and steroidogenic acute regulatory protein were lower. There were no differences in plasma cortisol concentration between the two charr populations immediately after capture and one hour after confinement. The strong biomarker response to OHCs and altered mRNA abundances of key genes related to HPI axis functioning in the Ellasjøen charr suggest endocrine disruptive effects of OHCs in this charr population. Possible ecological implications are not known, but it cannot be excluded that a slower growth rate in Ellasjøen charr compared to Laksvatn charr due to an increased metabolic demand associated with the activation of xenobiotic defense and detoxification systems may have contributed to the lower body mass of Ellasjøen charr compared to Laksvatn charr.

Fasting augments PCB impact on liver metabolism in anadromous arctic char.

Anadromous arctic char (Salvelinus alpinus) undertake short feeding migrations to seawater every summer and accumulate lipids, while the rest of the year is spent in fresh water where the accumulated lipid reserves are mobilized. We tested the hypothesis that winter fasting and the associated polychlorinated biphenyls' (PCBs) redistribution from lipid depots to critical tissues impair the liver metabolic capacity in these animals. Char were administered Aroclor 1254 (0, 1, 10, and 100 mg/kg body mass) orally and maintained for 4 months without feeding to mimic seasonal winter fasting, while fed groups (0 and 100 mg Aroclor 1254/kg) were maintained for comparison. A clear dose-related increase in PCB accumulation and cytochrome P4501A (CYP1A) protein content was observed in the livers of fasted fish. This PCB concentration and CYP1A response with the high dose of Aroclor were 1.5-fold and 3-fold greater in the fasted than in the fed fish, respectively. In fed fish, PCB exposure lowered liver glycogen content, whereas none of the other metabolic indicators were significantly affected. In fasted fish, PCB exposure depressed liver glycogen content and activities of glucose-6-phosphate dehydrogenase, alanine aminotransferase, lactate dehydrogenase, and phosphoenolpyruvate carboxykinase and elevated 3-hydroxyacylcoA dehydrogenase activity and glucocorticoid receptor protein expression. There were no significant impacts of PCB on heat shock protein 70 (hsp70) and hsp90 contents in either fed or fasted fish. Collectively, our study demonstrates that winter emaciation associated with the anadromous lifestyle predisposes arctic char to PCB impact on hepatic metabolism including disruption of the adaptive metabolic responses to extended fasting.