Osmoregulation in Atlantic salmon Salmo salar smolts transferred to seawater at different temperatures.

In order to investigate how changes in gill Na(+) , K(+) -ATPase (NKA) α1a and α1b subunits, Na(+) , K(+) , 2Cl(-) co-transporter (NKCC1) and the apical cystic fibrosis trans-membrane conductance regulator-I (CFTR-I) transcripts in wild strain of Atlantic salmon, Salmo salar, smolts are affected by temperature during spring, hatchery-reared parr (mean ± s.e. fork length = 14·1 ± 0·5; mean ± s.e. body mass = 28·5 ± 4·5 g) originating from broodstock from the Vosso river (western Norway) were acclimated to three temperature regimes (4·1, 8·1 and 12·9° C) in May and reared under gradually increasing salinity between May and June. Changes in plasma Na(+) , haematocrit (Hct) and PCO2 were monitored in order to assess and compare key physiological changes with the transcriptional changes in key ion transporters. The temperatures reflect the natural temperature range in the River Vosso during late spring. Overall, higher gill NKA α1b mRNA levels, gill NKCC1a levels and CFTR-I levels were observed in the 4·1° C group compared to the 11·9° C group. This coincided with a 2-3 week period with decreased Hct and PCO2 and may indicate a critical window when smolts suffer from reduced physical performance during migration. Further research is needed to confirm the potential interaction between ecological and physiological conditions on mortality of hatchery-reared smolts from River Vosso during their natural migration.

Pain and stress responses in farmed fish.

Farming fish for human consumption continues to expand as an industry and, with this increasing interaction with captive fish populations, there is now a growing interest in determining how to create good welfare for the fish we farm. This article summarises recent advances in our understanding of pain and stress responses in fish and how these relate to farmed fish welfare. Over the last decade several studies have examined whether or not fish feel pain, how aversive the experience is, and how such experiences may be mitigated through the use of analgesics. The basic neural mechanisms that enable the detection of tissue damage, i.e. nociceptive mechanisms, appear to be broadly conserved from fish through to birds and mammals, however, there is debate about the extent of the negative feelings associated with pain and whether these are truly experienced by fish. The stress response that helps fish to cope with various challenges also appears to be largely conserved across vertebrates, and the physiological changes that occur in response to acute and chronic stress in fish are similar to those described for mammals. Therefore, fish appear to have the innate ability to experience negative states such as pain and stress in a way analogous to that experienced by other vertebrates. There are multiple situations in which farmed fish may experience pain and stress and there is now a growing recognition that, to deliver appropriate welfare, on-farm practices and procedures will have to change. It is also the case that the welfare requirements of the different fish species that we farm vary, with some species coping better in captive rearing environments than others. The topic of fish welfare is relatively new and more research on stress responses, allostasis, pain thresholds and analgesics is required to help promote good fish welfare.

Aluminum exposure impacts brain plasticity and behavior in Atlantic salmon (Salmo salar).

Aluminum (Al) toxicity occurs frequently in natural aquatic ecosystems as a result of acid deposition and natural weathering processes. Detrimental effects of Al toxicity on aquatic organisms are well known and can have consequences for survival. Fish exposed to Al in low pH waters will experience physiological and neuroendocrine changes that disrupt homeostasis and alter behavior. To investigate the effects of Al exposure on both the brain and behavior, Atlantic salmon (Salmo salar) kept in water treated with Al (pH 5.7, 0.37±0.04 µmol 1(-1) Al) for 2 weeks were compared with fish kept in under control conditions (pH 6.7, <0.04 µmol 1(-1) Al). Fish exposed to Al and acidic conditions had increased Al accumulation in the gills and decreased gill Na(+), K(+)-ATPase activity, which impaired osmoregulatory capacity and caused physiological stress, indicated by elevated plasma cortisol and glucose levels. Here we show for the first time that exposure to Al in acidic conditions also impaired learning performance in a maze task. Al toxicity also reduced the expression of NeuroD1 transcript levels in the forebrain of exposed fish. As in mammals, these data show that exposure to chronic stress, such as acidified Al, can reduce neural plasticity during behavioral challenges in salmon, and may impair the ability to cope with new environments.

Seawater tolerance and post-smolt migration of wild Atlantic salmon Salmo salar × brown trout S. trutta hybrid smolts.

High levels of hybridization between Atlantic salmon Salmo salar and brown trout Salmo trutta have been reported in the River Driva. This study presents the underlying mechanisms of development of seawater (SW) tolerance and marine migration pattern for S. salar×S. trutta hybrids. Migrating S. salar×S. trutta hybrid smolts caught in the River Driva, Norway (a river containing Gyrodactylus salaris), displayed freshwater (FW) gill Na(+), K(+) -ATPase (NKA) activity levels of 11·8 µmol ADP mg protein h(-1), which were equal to or higher than activity levels observed in S. salar and S. trutta smolts. Following 4 days of SW exposure (salinity 32·3), enzyme activity remained high and plasma ion levels were maintained within the normal physiological range observed in S. salar smolts, indicating no signs of ion perturbations in S. salar×S. trutta hybrids. SW exposure induced an increase in NKA α1b-subunit mRNA levels with a concurrent decrease in α1a levels. Salmo salar×S. trutta post-smolts migrated rapidly through the fjord system, with increasing speed with distance from the river, as is often seen in S. salar smolts. The present findings suggest that S. salar×S. trutta smolts, as judged by the activity and transcription of the NKA system, regulation of plasma ion levels and migration speed more closely resemble S. salar than S. trutta.

Environmental effects on fish neural plasticity and cognition.

Most fishes experiencing challenging environments are able to adjust and adapt their physiology and behaviour to help them cope more effectively. Much of this flexibility is supported and influenced by cognition and neural plasticity. The understanding of fish cognition and the role played by different regions of the brain has improved significantly in recent years. Techniques such as lesioning, tract tracing and quantifying changes in gene expression help in mapping specialized brain areas. It is now recognized that the fish brain remains plastic throughout a fish's life and that it continues to be sensitive to environmental challenges. The early development of fish brains is shaped by experiences with the environment and this can promote positive and negative effects on both neural plasticity and cognitive ability. This review focuses on what is known about the interactions between the environment, the telencephalon and cognition. Examples are used from a diverse array of fish species, but there could be a lot to be gained by focusing research on neural plasticity and cognition in fishes for which there is already a wealth of knowledge relating to their physiology, behaviour and natural history, e.g. the Salmonidae.

Corticotropin-releasing factor neurogenesis during midlife development in salmon: genetic, environmental and thyroid hormone regulation.

Salmon parr-smolt transformation (smoltification) is a mid-life transitional stage between life in freshwater and seawater that entails a wide range of neural, endocrine and physiological modifications. In salmon, the neuroendocrine corticotropin-releasing factor (CRF) system regulates pituitary adrenocorticotrophic hormone and thyrotrophin release. Four experimental groups of Atlantic salmon, Salmo salar, were used to investigated CRF neurogenesis and its regulation during smoltification. We compared: (i) developmental stages (parr and early-smolt) in anadromous controls; (ii) a developmentally arrested model: anadromous reared under continuous light (LL) with anadromous controls; (iii) a natural hypoendocrine/incomplete smolt development salmon model (landlocked) with anadromous controls; and (iv) landlocked treated with thyroxine to anadromous control smolt levels. CRF neurogenesis between groups was studied with bromodeoxyuradine (BrdU) incorporation followed by double-labelling CRF and BrdU immunhistochemistry. The rate of CRF neurogenesis in the preoptic area (POA) increased from parr to early-smolts in anadromous salmon. By contrast, neurogenesis was inhibited in the LL group and reduced in the landlocked salmon. The administration of thyroxine in landlocked salmon to match anadromous levels increased the rate of CRF neurogenesis to anadromous levels. In conclusion, newly-formed CRF cells in the POA during smoltification are associated with increased retinal innervation to the POA and endocrine responsiveness to increased photoperiod. Both genetic and environmental factors influence the degree of salmon brain development. Thyroid hormones increase CRF neurogenesis during this critical period of development in salmon. We hypothesise that a positive-feedback of thyroid hormones on CRF neurogenesis may be an important event in reaching the developmental climax during critical periods.

Branchial expression patterns of claudin isoforms in Atlantic salmon during seawater acclimation and smoltification.

In euryhaline teleosts, permeability changes in gill epithelia are essential during acclimation to changed salinity. This study examined expression patterns of branchial tight junction proteins called claudins, which are important determinants of ion selectivity and general permeability in epithelia. We identified Atlantic salmon genes belonging to the claudin family by screening expressed sequence tag libraries available at NCBI, and classification was performed with the aid of maximum likelihood analysis. In gill libraries, five isoforms (10e, 27a, 28a, 28b, and 30) were present, and quantitative PCR analysis confirmed tissue-specific expression in gill when compared with kidney, intestine, heart, muscle, brain, and liver. Expression patterns during acclimation of freshwater salmon to seawater (SW) and during the smoltification process were examined. Acclimation to SW reduced the expression of claudin 27a and claudin 30 but had no overall effect on claudin 28a and claudin 28b. In contrast, SW induced a fourfold increase in expression of claudin 10e. In accord, a peak in branchial claudin 10e was observed during smoltification in May, coinciding with optimal SW tolerance. Smoltification induced no significant changes in expression of the other isoforms. This study demonstrates the expression of an array of salmon claudin isoforms and shows that SW acclimation involves inverse regulation, in the gill, of claudin 10e vs. claudin 27a and 30. It is possible that claudin 10e is an important component of cation selective channels, whereas reduction in claudin 27a and 30 may change permeability conditions in favor of the ion secretory mode of the SW gill.

Dopaminergic innervation of the rainbow trout pituitary and stimulatory effect of dopamine on growth hormone secretion in vitro.

To elucidate which factors regulate growth hormone (GH) secretion in rainbow trout, dopaminergic innervation of the rainbow trout pituitary along with the action of dopamine in vitro, were studied. Brains with attached pituitaries were double-labeled for putative dopaminergic neuronal fibers and somatotropes, using fluorescence immunohistochemistry. A direct dopaminergic innervation to the proximal pars distalis (PPD) with dopaminergic fibers terminating adjacent to somatotropes was demonstrated. Growth hormone secretion from whole pituitaries was measured in perifusate using a homologous GH-RIA. Dopamine (DA; 10(-7)-2x10(-6) g ml(-1)) increased basal GH secretion, with the GH secretion normalizing again after the DA exposure was halted. When pituitaries were pre-treated with somatostatin-14 (SRIF-14; 10(-12)-10(-9) g ml(-1)), before being exposed to different doses of DA, there was an inhibition of GH secretion which was not reversed after treatment of SRIF-14 was halted, unless DA was added. It is concluded that dopamine can function as a GH secretagogue in the rainbow trout pituitary gland.

Diabetes is related to fatty acid imbalance in Eskimos.

The reasons for the rapidly increasing prevalence of diabetes (NIDDM) among Alaskan Eskimos are only partly understood. This study examines the association of fatty acid metabolism in 68 Alaskan Eskimos with NIDDM or impaired glucose tolerance (IGT) and 386 with normal glucose tolerance > 24 years old. The prevalence of NIDDM was 12% and IGT was 18% in those > 54 years of age and in those < 55 years of age was 3.7% and 3.0%, respectively. Those with abnormal glucose tolerance had lower concentrations of some omega-3 fatty acids (FAs 18:3 omega-3, 20:5 omega-3) and some omega-6 FAs (18:3 omega-6, 20:3 omega-6, 22:4 omega-6) and higher concentrations of palmitic acid (16:0) and oleic acid (18:1 omega-9) than the normo-glycemic participants. These data provide evidence that glucose intolerance and insulin resistance are associated with a deviation from a traditional diet of fish and marine mammals (high in omega-3 FAs and low in saturated fats) to commercial foods (low in omega-3 FAs and high in saturated fats). The low plasma concentrations of the long-chain omega-6 FAs in the glucose impaired may reflect a defect in desaturase activity.

Diabetes and impaired glucose tolerance in three Alaskan Eskimo populations. The Alaska-Siberia Project.

OBJECTIVE: The objectives of this study were to determine the prevalence of diabetes and impaired glucose tolerance (IGT) in three Alaskan Eskimo populations, using standardized diagnostic criteria, and to evaluate family history and obesity as risk factors. RESEARCH DESIGN AND METHODS: This cross-sectional study involved men and women > or = 25 years of age from three Eskimo ethnic groups (Siberian Yupik, Central Yupik, and Inupiat) residing in northwestern Alaska. Glucose tolerance status was defined by World Health Organization criteria and was based on a 75-g oral glucose tolerance test. Data on age, family history of diabetes, and degree of Eskimo ancestry were obtained from a personal interview. Obesity was assessed using BMI. RESULTS: A total of 454 of 899 (50.5%) eligible participants were examined for diabetic status (239 Siberian Yupik, 106 Central Yupik, and 109 Inupiat participants). The prevalence of diabetes was more than twice as high among the Siberian Yupik (9.6%) as among the Central Yupik (2.8%) and Inupiat participants (3.7%). Diabetes was more prevalent in women than men (8.8 vs. 4.2%). IGT was found in an additional 11.7% of the women and 4.7% of the men. The combined prevalence of diabetes and IGT in the population > or = 55 years of age was 30.4% (diabetes 12.0%, IGT 18.4%). Of the people identified with diabetes, 47% had not been previously diagnosed. Age-specific prevalences were similar to those found in U.S. whites in the National Health and Nutrition Examination Survey II. After adjustment for age, family history of diabetes was associated with diabetes in study participants with an odds ratio of 4.4, while obesity was associated with diabetes with an odds ratio of 2.6. CONCLUSIONS: These prevalences of diabetes are the highest yet reported among Eskimo populations. Obesity and family history of diabetes are associated with increased odds of developing diabetes. These data underscore the need to further examine risk factors and to design effective interventions.

Transient alterations in neurotransmitter levels during a critical period of neural development in coho salmon (Oncorhyncus kisutch).

The time of parr-smolt transformation (PST) in salmon is a critical period of brain development during which the olfactory imprinting on the natal stream takes place. PST is associated with a surge of plasma thyroxine. We report here similar surges in the brain content of serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), dopamine (DA), norepinephrine (NE), and glutamine (Gln). Other putative neurotransmitters, such as glutamate and aspartate show no such change while gamma-amino-butyric acid (GABA) shows a minor elevation.

Lipoprotein profiles in Alaskan Siberian Yupik Eskimos.

Coronary heart disease and diabetes, once rare in Eskimos, is on the increase in some Alaskan communities. As part of a detailed assessment of the prevalence of these diseases and associated risk factors in several villages, we report here on the plasma concentrations of lipoprotein and apoprotein in a sample of Siberian Yupik Eskimos aged 40-87 years living in the village of Gambell on St. Lawrence Island. Mean cholesterol levels for females were 242 mg/dl and 223 for males. LDL levels were 161 for females and 149 for males, while HDL levels were 67 for females and 58 for males. The mean ApoB and Apo-A1 values were 112 mg/dl and 167 mg/dl for males and females. Triglycerides were 73 for females and 77 for males. The allele frequency of APOE*3 and APOE*4 were .900 and .100 respectively. There was a total absence of the APOE*2 allele in this sample. Mean total cholesterol concentrations in this sample were markedly higher than those reported in 1958 from this village and from those recently reported for closely related Yupik Eskimos living in Siberia. The cholesterol levels were higher and the triglyceride levels were lower than in U.S. Indian populations. The data suggest the possibility of recent increased risk of cardiovascular disease for this Eskimo population. The new information indicates a need for comprehensive epidemiological studies to identify and characterize cardiovascular disease risk factors in all Alaska Native populations in order to provide a database for meaningful interventions. The lipoprotein profiles reported here are significantly different from Amerind groups, a finding that may reflect both dietary and genetic differences.

Distribution and changes in mu- and kappa-opiate receptors during the midlife neurodevelopmental period of Coho salmon, Oncorhynchus kisutch.

Parr-smolt transformation (PST) in coho salmon is associated with a plasma thyroid hormone (PT4) surge and a critical period of neural development that includes axonal sprouting, neurogenesis, and surges of select neurotransmitters. Here we provide a description of the selectivity, distribution, and the changes in the density of mu- and kappa-opiate receptors during PST, as revealed by quantitative in vitro autoradiography of [3H]Tyr-D-Ala-Gly-NMe-Phe-Gly-ol ([3H]DAMGO) and [3H]ethylketocyclazocine ([3H]EKC), respectively. The concentration of mu-receptors increased significantly in select cell groups in the early stages of parr-smolt transformation, until a peak was reached at the time coinciding with the peak of the PT4 surge. In other cell groups, the peaks occurred 1 or 2 weeks later. With one exception, this increase was followed by a decrease in concentration. The brain areas showing the highest concentrations are the dorsal nucleus of the ventral telencephalic area, the glomerular region, the granular layer of the valvula cerebelli, the nucleus diffuses of the inferior lobe, and the nucleus diffuses of the torus lateralis. Other regions with distinctly elevated mu-receptor concentrations are the stratum griseum centrale of the optic tectum and the preoptic area. The distribution of kappa-receptors is more diffuse, and the densities are considerably lower. The overlap in distribution of mu- and kappa-receptors is considerable, but significant exceptions are noted. For example, the dorsomedial nucleus of the dorsal telencephalic area, the habenular nucleus, and the dorsomedial nucleus of the thalamus exhibit a surge in density of kappa-receptors at the time of the PT4 surge, while the density of mu-receptors in these nuclei remain very low throughout parr-smolt transformation. The kappa-receptor containing cell groups are not identifiable until 3 weeks before the PT4 surge because of low densities. The most prominently labeled kappa-receptor regions are the ventral and dorsal nuclei of the ventral telencephalic area, the medial and dorsal zones of the dorsal telencephalic area, the optic tectum (all layers), the dorsomedial nucleus of the thalamus, the torus lateralis of the ventral hypothalamus, and the preoptic area. An increase of mu- and kappa-opiate receptor densities in specific brain regions may reflect roles in the alteration of brain organization, olfactory imprinting, neuroendocrine activity or other physiological activities. The overall distribution of these receptors are relatively more extensive in salmon than in other vertebrates so far studied.

Transient serotonin-immunoreactive neurons coincide with a critical period of neural development in coho salmon (Oncorhynchus kisutch).

In coho salmon (Oncorhynchus kisutch), smolt transformation has been shown to be associated with sequential surges of neurotransmitters in the brain. In order to determine if the surge of serotonin (5-HT) is correlated with structural changes, we have used immunocytochemistry to observe changes in the serotonin immunoreactivity before, during and after the 5-HT surge. The following stages were studied: 12-month-old freshwater presmolts, 17-month-old freshwater presmolts, 18-month-old saltwater smolts, 19-month-old saltwater postsmolt, 24-month-old postsmolt, and adult spawners. In the 17-month-old samples, but not at any other stage, we found a set of transient (serotonin-immunoreactive) 5-HT-immunoreactive neurons in the lateral preoptic area, as well as a discrete population of 5-HT-immunoreactive neurons in the lateral part of the dorsal right habenular nucleus. In addition, a higher density of serotonergic fibers was found in the telencephalon at this stage compared to the following two stages. Since the transient 5-HT-immunoreactive structures presented here do not appear simultaneously with the 5-HT total brain concentration surge, we conclude that they are unlikely to be the source of the 5-HT surge, but are probably related to other developmental changes in the brain associated with smolt transformation.

Pseudomonas cepacia 2,2-dialkylglycine decarboxylase. Sequence and expression in Escherichia coli of structural and repressor genes.

A 3969-base pair PstI-PstI fragment of Pseudomonas cepacia DNA containing the gene for the pyridoxal 5'-phosphate dependent 2,2-dialkylglycine decarboxylase (pyruvate) (EC 4.1.1.64) was cloned in Escherichia coli. The insert was sequenced by the dideoxy method using nested deletions from both ends, revealing a central 1302-base pair region that codes for the decarboxylase subunit. The recombinant enzyme was expressed in E. coli, purified to homogeneity, and sequenced at the amino terminus. Also, a cofactor-labeled active site peptide was sequenced. The carboxyl terminus of the deduced amino acid sequence is homologous with the carboxyl terminus of mammalian ornithine aminotransferase; the active site sequence is similar to the active site sequences of several other aminotransferases. No homologies with known decarboxylase sequences could be found. Expression of the decarboxylase gene is negatively controlled by a 687-nucleotide sequence upstream of and diverging from the structural gene. Expression is induced by S-isovaline, 2-methylalanine, and D-2-aminobutanoic acid, but not by glycine, D- or L-alanine, L-2-aminobutanoic acid, R-isovaline, or other alkyl amino acids.