Seawater Challenge Research Articles

Crosstalk between Growth and Osmoregulation of GHRH-SST-GH-IGF Axis in Triploid Rainbow Trout (Oncorhynchus mykiss).

Smolting is an important development stage of salmonid, and an energy trade-off occurs between osmotic regulation and growth during smolting in rainbow trout (Oncorhynchus mykiss). Growth hormone releasing hormone, somatostatin, growth hormone and insulin-like growth factor (GHRH-SST-GH-IGF) axis exhibit pleiotropic effects in regulating growth and osmotic adaptation. Due to salmonid specific genome duplication, increased paralogs are identified in the ghrh-sst-gh-igf axis, however, their physiology in modulating osmoregulation has yet to be investigated. In this study, seven sst genes (sst1a, sst1b, sst2, sst3a, sst3b, sst5, sst6) were identified in trout. We further investigated the ghrh-sst-gh-igf axis of diploid and triploid trout in response to seawater challenge. Kidney sst (sst1b, sst2, sst5) and sstr (sstr1b1, sstr5a, sstr5b) expressions were changed (more than 2-fold increase (except for sstr5a with 1.99-fold increase) or less than 0.5-fold decrease) due to osmoregulation, suggesting a pleiotropic physiology of SSTs in modulating growth and smoltification. Triploid trout showed significantly down-regulated brain sstr1b1 and igfbp2a1 (p < 0.05), while diploid trout showed up-regulated brain igfbp1a1 (~2.61-fold, p = 0.057) and igfbp2a subtypes (~1.38-fold, p < 0.05), suggesting triploid trout exhibited a better acclimation to the seawater environment. The triploid trout showed up-regulated kidney igfbp5a subtypes (~6.62 and 7.25-fold, p = 0.099 and 0.078) and significantly down-regulated igfbp5b2 (~0.37-fold, p < 0.05), showing a conserved physiology of teleost IGFBP5a in regulating osmoregulation. The IGFBP6 subtypes are involved in energy and nutritional regulation. Distinctive igfbp6 subtypes patterns (p < 0.05) potentially indicated trout triggered energy redistribution in brain and kidney during osmoregulatory regulation. In conclusion, we showed that the GHRH-SST-GH-IGF axis exhibited pleiotropic effects in regulating growth and osmoregulatory regulation during trout smolting, which might provide new insights into seawater aquaculture of salmonid species.

Dietary effects on biomarkers of growth, stress, and welfare of diploid and triploid Atlantic salmon (Salmo salar) during parr-smolt transformation

Triploidy is induced in Atlantic salmon (Salmo salar) to produce sterile fish for genetic containment and to hinder early sexual maturation in farmed fish, but it can have unwanted negative effects on growth, health, and welfare. However, the growth and welfare of triploid fish may be improved by adjusting the rearing environment, feeding conditions and diets. This study evaluated physiological changes and used a suite of biomarkers to assess the potential impact of diet on growth and welfare of diploid and triploid salmon during the parr-smolt transformation. Diploids and triploids, held at low temperature, were fed a standard salmon feed or one with hydrolyzed fish proteins thought to be suitable for triploid Atlantic salmon. Fish muscle was collected monthly from October to December (2454–3044 degree-days post-start feeding, ddPSF) for analysis of biomarkers, and the progress of the parr-smolt transformation was monitored using a seawater challenge test. Real-Time PCR and radioimmunoassay were used to assess growth and stress response biomarkers (expression of genes of the GH-IGF axis and HSP70; cortisol concentrations), and oxidative stress biomarkers of lipids (MDA) and proteins (AOPP) were assayed. Changes in the biomarkers were related to sampling time rather than being associated with diet or ploidy, and the changes were compatible with the progression of the parr-smolt transformation. Growth and expressions of the biomarkers in triploid Atlantic salmon were similar to those of their diploid counterparts, and there was no evidence that the rearing conditions employed in the study resulted in stress responses being elicited. Overall, the physiological indicators and biomarkers employed in this study did not point to there being any dietary effects on performance and welfare of diploid and triploid salmon that were undergoing parr-smolt transformation.

Establishment of a Simplified System to Evaluate Salinity Preference and Validation of Behavioral Salinity Selection in the Japanese Medaka, Oryzias latipes

In fishes, it is necessary to select a salinity environment suitable for survival. However, little is known about the mechanisms regarding detection and selection of salinity environments in fish. This study involved the establishment of a simple aquarium system in which fish can swim between freshwater (FW) and seawater (SW) in a single tank. In this tank, the lower level contained SW, the upper level contained FW, and the FW and SW levels were clearly separated as different salinity areas. Behavioral experiments of salinity environment selection using this simplified system to evaluate salinity preference showed that FW-acclimated medakas preferred FW to SW. In contrast, SW-acclimated medakas preferred SW to FW. These results indicate that euryhaline medakas prefer the saline habitats to which they are acclimated, when able to select the salinity environment. We identified the taste receptor type-2 and polycystic kidney disease 2-like 1 genes as possibly related to high-salinity taste in medaka. The expression of these genes increased at certain time points after SW challenges. In this study, we established an aquarium system to facilitate a simple experiment for salinity preference. Our results suggest that the medaka is good model for research related to seawater environment selection in fish.

Corticosteroid control of Na+/K+-ATPase in the intestine of the sea lamprey (Petromyzon marinus)

Anadromous sea lamprey (Petromyzon marinus) larvae undergo a months-long true metamorphosis during which they develop seawater (SW) tolerance prior to downstream migration and SW entry. We have previously shown that intestinal Na+/K+-ATPase (NKA) activity increases during metamorphosis and is critical to the osmoregulatory function of the intestine in SW. The present study investigated the role of 11-deoxycortisol (S) in controlling NKA in the anterior (AI) and posterior (PI) intestine during sea lamprey metamorphosis. In a tissue profile, nka mRNA and protein were most abundant in the gill, kidney, and AI. During metamorphosis, AI nka mRNA increased 10-fold, whereas PI nka mRNA did not change. Specific corticosteroid receptors were found in the AI, which had a higher binding affinity for S compared to 11-deoxycorticosterone (DOC). In vivo administration of S in mid-metamorphic lamprey upregulated NKA activity 3-fold in the AI and PI, whereas administration of DOC did not affect intestinal NKA activity. During a 24 h SW challenge test, dehydration of white muscle moisture was rescued by prior treatment with S, which was associated with increased intestinal nka mRNA and NKA activity. These results indicate that intestinal osmoregulation in sea lamprey is a target for control by S during metamorphosis and the development of SW tolerance.

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.

Effects of seawater acclimation at constant and diel cyclic temperatures on growth, osmoregulation and branchial phospholipid fatty acid composition in rainbow trout Oncorhynchus mykiss.

The study investigated the effects of seawater acclimation at constant and diel temperatures on the growth, osmoregulation, and branchial phospholipid fatty acid (PLFA) composition in rainbow trout (Oncorhynchus mykiss). The fish (initial weight, 62.28 ± 0.41g) were reared at a constant 13.0°C (CT) or with a diel cycle of either 13.0 ± 1.0°C (VT2) or 13.0 ± 2.0°C (VT4) for 6weeks, and subsequently subjected to seawater acclimation. Diel temperature variations (of up to 4°C) did not affect the growth rate of rainbow trout maintained in freshwater, but alleviated the impairment on the growth after seawater challenge. Under all temperature conditions, rainbow trout were well prepared to seawater acclimation. The diel cyclic temperature resulted in fish with reduced fluctuations in plasma electrolyte levels, branchial Na+-K+ ATPase activity, and plasma osmolality. In freshwater, the sum of the monounsaturated fatty acids was significantly higher in the VT4 relative to CT and VT2 treatment. Conversely, the sum of polyunsaturated fatty acids was significantly lower in the VT4 fish. After seawater transfer, the branchial PLFA profiles of the fish significantly changed, but those in CT and VT2 did not recover afterwards (the degree of unsaturation was downregulated). The PLFA composition of fish in the VT4 treatment appeared to be steadier under seawater acclimation. This study suggests that a diel cyclic temperature (13.0 ± 2.0°C) can alleviate the impairment of growth, enhance osmoregulation capability, and improve the stability of the branchial PLFA composition in rainbow trout after seawater acclimation.

Differential Seawater Adaptability in Three Different Sizes of Under-yearling Steelhead Trout.

Seawater adaptability of steelhead trout increases along with the increase in the size of the fish, independent of parr-smolt transformation. Three 96 h seawater challenge tests were conducted to determine the size at which seawater adaptability of steelhead trout develops. Plasma Na+ and Cl- levels, moisture content, gill Na+/K+ ATPase activity, and mortality during the 96 h after direct transfer to seawater (32 ppt) were determined. Plasma Na+ and Cl- levels in 50 g fish continuously increased during the 96 h after the transfer to seawater (p<0.05), but the levels in 100 and 150 g fish leveled off after 24 h (p<0.05). Both 100 and 150 g size steelhead trout maintained muscle moisture content (%) better than 50 g size fish (p<0.05). Gill Na+/K+ ATPase activity in the 100 g size group increased in a time-dependent manner after transfer to seawater (p<0.05), whereas activity in the 50 and 150 g sizes did not increase (p>0.05), for which a possible explanation was discussed. A mere 2.6% mortality in both the 50 and 150 g size groups was observed. In conclusion, the current results indicate that 50 g size steelhead trout did not show development of a high level of hypoosmoregulatory capacity, whereas fish in the 100 and 150 g size groups showed a high level in our experimental conditions. Therefore, the steelhead trout larger than a 100 g size is recommended for transfer to seawater culture.

Insulin-like growth factor 1 triggers salt secretion machinery in fish under acute salinity stress

Timely adjustment of osmoregulation upon acute salinity stress is essential for the survival of euryhaline fish. This rapid response is thought to be tightly controlled by hormones; however, there are still questions unanswered. In this work, we tested the hypothesis that the endocrine hormone, insulin-like growth factor 1 (Igf1), a slow-acting hormone, is involved in the activation of salt secretion mechanisms in euryhaline medaka (Oryzias melastigma) during acclimation to acute salinity stress. In response to a 30-ppt seawater (SW) challenge, Na+/Cl- secretion was enhanced within 0.5 h, with concomitant organization of ionocyte multicellular complexes and without changes in expression of major transporters. Igf1 receptor inhibitors significantly impair the Na+/Cl- secretion and ionocyte multicellular complex responses without affecting transporter expression. Thus, Igf1 may activate salt secretion as part of the teleost response to acute salinity stress by exerting effects on transporter function and enhancing the formation of ionocyte multicellular complexes. These findings provide new insights into hormonal control of body fluid ionic/osmotic homeostasis during vertebrate evolution.

Diminished Expression of Na+/HCO3−cotransporter 1.2a (nbce1.2a) mRNA Accompanies Seawater Acclimation in Atlantic Salmon

The life history of anadromous Atlantic salmon (Salmo salar) entails an initial freshwater stage that precedes their migration to marine environments. The development of physiological systems that will ultimately support the survival of juvenile salmon upon their entry into marine habitats is termed parr‐smolt transformation. This springtime transformation (smoltification) includes the acquisition of seawater (SW) tolerance. Thus, the ability of smolts to successfully acclimate to SW is attained through the development of branchial pathways for ion‐secretion. In addition, the capacity to rapidly attenuate ion‐absorptive processes upon exposure to SW is a vital aspect of SW tolerance. In the current study, we investigated the branchial gene expression patterns of three Na+/HCO3−cotransporter 1 isoforms (nbce1.1, nbce1.2a, and nbce1.2b) during smoltification and following seawater challenges. Given the proposed roles of Nbce1s in the absorption of environmental Na+ by branchial ionocytes in other teleosts, we hypothesized that nbce1s would be downregulated in salmon undergoing smoltification and/or following SW exposure. In two separate years, we observed springtime increases in branchial Na+/K+‐ATPase activity, Na+/K+/2Cl−‐cotransporter 1 and cystic fibrosis transmembrane regulator 1 expression characteristic of smoltification, whereas there were no seasonal changes in the expression of nbce1 isoforms. On the other hand, we observed diminished nbce1.2a expression within 2 days after the transfer of parr and smolts to SW that remained low throughout the experiment (&gt;2 weeks). Collectively, our results suggest that while there are no clear seasonal nbce1 dynamics during smoltification, the modulation of nbce1.2a expression upon exposure to SW may underlie the attenuation of environmental Na+ uptake.Support or Funding InformationSupported by the National Science Foundation (IOS‐1755131).

A temperature shift on the migratory route similarly impairs hypo-osmoregulatory capacities in two strains of Atlantic salmon (Salmo salar L.) smolts

Anthropogenic use of water systems may cause temperature fluctuations between tributaries and large rivers for which physiological population related-effects on osmoregulatory capacity of Atlantic salmon are not well described. We simulated the downstream route in the case of the River Meuse basin to investigate the impact of a 5°C temperature shift during smoltification on hypo-osmoregulatory capacities of smolts. Three temperature regimes were tested: control temperature-treatment (T1) without temperature shift, early (T2) or late (T3) temperature shift-treatment. Moreover, fish were subjected to seawater challenge during and after the downstream migration peak time. Two allochtonous strains were used: Loire-Allier (LA) and Cong (CG). Without temperature shift (T1), significant differences between the strains were noticed in the peak date and maximum activity of gill Na+/K+ATPase as well as in plasma sodium and potassium concentrations. For early (T2) and late (T3) temperature shift-treatments, gill Na+/K+ATPase activity, plasma osmolality and ion concentrations were negatively influenced in both strains. After salinity challenge, the highest osmolality was measured in smolts subjected to the temperature shift. Predictably circulating levels of GH and IGF-1 changed over the smolting period but they did not explain the observed modifications in hypo-osmoregulatory abilities whatever the population. The results show a negative impact of a temperature shift on hypo-osmoregulatory capacities of smolts regardless of population differences in smoltification timing under conditions without temperature shift. The resilience of such physiological impact was sustained at least for 1week, comforting the role of high temperature in influencing the rate of changes occurring during smoltification. Therefore, favouring the downstream migration to help smolts reach the sea faster may mitigate the impact of a rapid temperature increase.

Does silvering or 11-ketotestosterone affect osmoregulatory ability in the New Zealand short-finned eel (Anguilla australis)?

Silvering has been associated with advancing osmoregulatory ability. Given the demonstrated role of 11-ketotestosterone (11KT) in mediating many of the silvering-related changes, we investigated the role of 11KT in driving this advanced osmoregulatory ability in the New Zealand short-finned eel (Anguilla australis). Yellow (non-migratory) eels with or without 11KT implants and blank-implanted silver (migratory) eels, either held in freshwater or subjected to seawater challenge, were sampled to determine serum [Na+] and [Cl-], pituitary prolactin mRNA levels, gill Na+/K+-ATPase activity and gill mRNA levels for Na+/K+-ATPase-α1 subunit and for Na+/K+/2Cl- co-transporter-1α-subunit. Developmental stage and 11KT treatment advanced the eels' osmoregulatory ability. Thus, serum [Na+] and [Cl-] were affected by developmental stage and 11KT treatment upon seawater challenge. However, seawater challenge, not 11KT treatment or developmental stage, produced the strongest and the most consistent effects on A. australis osmoregulatory processes, inducing significant effects in all the relevant parameters we measured. In light of our results and in view of the eel's marine ancestry, we contend that A. australis, or freshwater eels in general, are highly tolerant and able to adapt quickly to changing salinities even at the yellow stage, which may preclude a critical need for an advanced osmoregulatory ability at silvering.

Rearing strategies alter patterns of size-selective mortality and heritable size variation in steelhead trout (Oncorhynchus mykiss)

Steelhead trout (Oncorhynchus mykiss) reared under two different regimes: high food ration for 1 year (S1; typical strategy) or low ration for 2 years (S2) were subjected to a seawater challenge during the corresponding spring outmigration period. The S1smolts were smaller and suffered greater seawater challenge mortality (23.9% compared with 0.7% for the S2smolts) that was significantly and negatively related to body size. Heritability of body size was similar for the two treatments during the parr stage (fork length: S1= 0.181, S2= 0.245; mass: S1= 0.372; S2= 0.447), but higher for the S1treatment during the smolt stage for length (S1= 0.212, S2= 0.002) and body mass (S1= 0.145, S2= 0.015). Strong family effects for both traits and significant family by environment interactions for parr mass and smolt length indicated significant phenotypic plasticity. A genetic response to size-selective mortality caused by insufficient growth opportunity in the S1treatment is plausible and may affect fitness in the natural environment through effects on correlated traits.

Variation in branchial expression among insulin-like growth-factor binding proteins (igfbps) during Atlantic salmon smoltification and seawater exposure

BackgroundIn preparation for migration from freshwater to marine habitats, Atlantic salmon (Salmo salar L.) undergo smoltification, a transformation that includes the acquisition of hyposmoregulatory capacity. The growth hormone (Gh)/insulin-like growth-factor (Igf) axis promotes the development of branchial ionoregulatory functions that underlie ion secretion. Igfs interact with a suite of Igf binding proteins (Igfbps) that modulate hormone activity. In Atlantic salmon smolts, igfbp4,−5a,−5b1,−5b2,−6b1 and−6b2 transcripts are highly expressed in gill. We measured mRNA levels of branchial and hepatic igfbps during smoltification (March, April, and May), desmoltification (July) and following seawater (SW) exposure in March and May. We also characterized parallel changes in a broad suite of osmoregulatory (branchial Na+/K+-ATPase (Nka) activity, Na+/K+/2Cl−cotransporter 1 (nkcc1) and cystic fibrosis transmembrane regulator 1 (cftr1) transcription) and endocrine (plasma Gh and Igf1) parameters.ResultsIndicative of smoltification, we observed increased branchial Nka activity, nkcc1 and cftr1 transcription in May. Branchial igfbp6b1 and -6b2 expression increased coincidentally with smoltification. Following a SW challenge in March, igfbp6b1 showed increased expression while igfbp6b2 exhibited diminished expression. igfbp5a,−5b1 and−5b2 mRNA levels did not change during smolting, but each had lower levels following a SW exposure in March.ConclusionsSalmonids express an especially large suite of igfbps. Our data suggest that dynamic expression of particular igfbps accompanies smoltification and SW challenges; thus, transcriptional control of igfbps may provide a mechanism for the local modulation of Igf activity in salmon gill.

Genetic basis for variation in salinity tolerance between stickleback ecotypes.

Adaptation to different salinities can drive and maintain divergence between populations of aquatic organisms. Anadromous and stream ecotypes of threespine stickleback (Gasterosteus aculeatus) are an excellent model to explore the genetic mechanisms underlying osmoregulation divergence. Using a parapatric pair of anadromous and stream stickleback ecotypes, we employed an integrated genomic approach to identify candidate genes important for adaptation to different salinity environments. Quantitative trait loci (QTL) mapping of plasma sodium concentrations under a seawater challenge experiment identified a significant QTL on chromosome 16. To identify candidate genes within this QTL, we first conducted RNA-seq and microarray analysis on gill tissue to find ecotypic differences in gene expression that were associated with plasma Na+ levels. This resulted in the identification of ten candidate genes. Quantitative PCR analysis on gill tissue of additional Japanese stickleback populations revealed that the majority of the candidate genes showed parallel divergence in expression levels. Second, we conducted whole-genome sequencing and found five genes that are predicted to have functionally important amino acid substitutions. Finally, we conducted genome scan analysis and found that eight of these candidate genes were located in genomic islands of high differentiation, suggesting that they may be under divergent selection. The candidate genes included those involved in ATP synthesis and hormonal signalling, whose expression or amino acid changes may underlie the variation in salinity tolerance. Further functional molecular analysis of these genes will reveal the causative genetic and genomic changes underlying divergent adaptation.

Passing a seawater challenge test is not indicative of hatchery-reared Atlantic salmon Salmo salar smolts performing as well at sea as their naturally produced conspecifics.

Despite satisfactory reactions to seawater challenge tests indicative of appropriate physiological state, hatchery-reared Atlantic salmon Salmo salar smolts stocked in the Eira River in Norway between 2001 and 2011 performed less well at sea in terms of growth, age at maturity and survival than smolts of natural origin. The mean rates of return to the river for hatchery-reared and naturally produced S. salar were 0·98 and 2·35%. In the Eira River, c. 50 000 hatchery-reared S. salar smolts of local origin were stocked annually to compensate for reduced natural smolt production following regulation for hydroelectric purposes, while a mean of 17 262 smolts were produced naturally in the river. This study demonstrates that, although captive S. salar perform well in seawater challenge tests, hatchery-reared smolts are not necessarily as adaptable to marine life as their naturally produced counterparts. These findings suggest that production of hatchery-reared smolts more similar to naturally produced individuals in morphology, physiology and behaviour will be necessary to improve success of hatchery releases. Where possible, supplementary or alternative measures, including habitat restoration, could be implemented to ensure the long-term viability of wild stocks.

Latent Toxicity of Endothall to Anadromous Salmonids During Seawater Challenge

Limited evidence exists on the latent effects of toxicant exposure on the seawater adaptability of anadromous salmon and steelhead. It is unclear whether such an effect exists for the widely used and relatively non-toxic herbicide endothall. Coho salmon, Oncorhynchus kisutch (coho), Chinook salmon, O. tshawytscha (Chinook), and anadromous rainbow trout, O. mykiss (steelhead) were subjected to a 10-day seawater challenge following freshwater treatments [0-12 mg acid equivalent (a.e)./L at 96 h]. Mean survival resulted in 82 % (n = 225), 84 % (n = 133), 90 % (n = 73) and 59 % (n = 147) survival for 0, 3-5, 6-8, and 9-12 mg a.e./L, respectively. Our results indicate a lower toxicity threshold compared with previously reported acute toxicity results, but higher compared with previous seawater challenge studies. We demonstrate the utility of the seawater challenge assay to accurately define toxic effects of pesticides on salmonids with complex life-histories.

Activation of TRPV1-dependent calcium oscillation exacerbates seawater inhalation-induced acute lung injury.

Calcium is an important second messenger and it is widely recognized that acute lung injury (ALI) is often caused by oscillations of cytosolic free Ca2+. Previous studies have indicated that the activation of transient receptor potential-vanilloid (TRPV) channels and subsequent Ca2+ entry initiates an acute calcium-dependent permeability increase during ALI. However, whether seawater exposure induces such an effect through the activation of TRPV channels remains unknown. In the current study, the effect of calcium, a component of seawater, on the inflammatory reactions that occur during seawater drowning-induced ALI, was examined. The results demonstrated that a high concentration of calcium ions in seawater increased lung tissue myeloperoxidase activity and the secretion of inflammatory mediators, such as tumor necrosis factor-α (TNF-α) and interleukin (IL)-1β and IL-6. Further study demonstrated that the seawater challenge elevated cytosolic Ca2+ concentration, indicated by [Ca2+]c, by inducing calcium influx from the extracellular medium via TRPV1 channels. The elevated [Ca2+c] may have resulted in the increased release of TNF-α and IL-1β via increased phosphorylation of nuclear factor-κB (NF-κB). It was concluded that a high concentration of calcium in seawater exacerbated lung injury, and TRPV1 channels were notable mediators of the calcium increase initiated by the seawater challenge. Calcium influx through TRPV1 may have led to greater phosphorylation of NF-κB and increased release of TNF-α and IL-1β.

Variation in Branchial Expression Among insulin‐like growth‐factor binding proteins (igfbps) During Atlantic Salmon Smoltification

In preparation for migration from freshwater to marine habitats, Atlantic salmon (Salmo salar) undergo smoltification, a springtime transformation that includes the development of salinity tolerance. The growth hormone/insulin‐like growth‐factor (Igf) network is known to mediate the development of branchial ionoregulatory functions underlying salinity tolerance. Igfs interact with a suite of Igf binding proteins (Igfbps) that modulate hormone activity. In Atlantic salmon, igfbp4, ‐5a, ‐5b1, ‐5b2, ‐6b2 and ‐6b1 are highly expressed in branchial tissue. Here, we characterized the dynamics of igfbp expression during smoltification and following acute seawater (SW) challenges. We observed increased branchial Na+, K+‐ATPase activity and Na+,K+,2Cl‐‐cotransporter expression characteristic of smoltification. igfbp5a, ‐5b1 and ‐5b2 expression did not peak in Spring. Nonetheless, igfbp5a, ‐5b1 and ‐5b2 showed diminished expression following SW challenge in March. igfbp6b1 and ‐6b2 expression, however, did increase in Spring, peaking with smoltification. Following a SW challenge in March, igfbp6b1 showed increased expression while igfbp6b2 exhibited diminished expression. These data suggest that dynamic igfbp expression accompanies smoltification and SW challenges. Transcriptional control of igfbps may thus provide a mechanism for the local modulation of Igf activity in gill.

Seasonal changes in gene expression of corticoid receptors in anadromous and non-anadromous strains of rainbow trout Oncorhynchus mykiss.

To clarify the regulation of expression of corticoid receptor (CR) genes during period of parr-smolt transformation of salmonids, seasonal changes in mRNA levels of glucocorticoid receptor (GR)-1, GR-2 and mineralocorticoid receptor (MR) were examined in gill, leucocytes, spleen and brain of anadromous and non-anadromous forms of Oncorhynchus mykiss. Increases in gill Na(+) , K(+) ATPase activity, plasma thyroxine levels and hypo-osmoregulatory ability assessed by 24 h seawater challenge test represented characteristics of smoltification in anadromous O. mykiss from May to June, whereas there was no apparent increase in the values of non-anadromous O. mykiss. Plasma cortisol levels of anadromous O. mykiss were higher than levels of non-anadromous O. mykiss from April to June. In gill of non-anadromous O. mykiss, there were significant increases in mRNA levels of three types of CR in spring. Although there were significant seasonal variations of CR mRNA levels in gill of anadromous O. mykiss, they appear to be less clear than those variations in non-anadromous O. mykiss. In anadromous O. mykiss, significant elevations in mRNA levels of the three types of CR were observed especially in the spleen. In both preoptic area and basal hypothalamus of the brain, there were tendencies to increase in CR mRNA levels from spring to summer in both anadromous and non-anadromous O. mykiss. These results showed difference in regulation of CR gene expression between the two forms of O. mykiss for osmoregulatory, immune and central nervous systems.

Long‐term hypo‐osmoregulatory capacity in downstream migrating Atlantic salmon Salmo salar L. smolts

The duration of hypo-osmoregulatory capacity in downstream migrating Atlantic salmon Salmo salar L smolts previously stocked as startfed young-of-the year (YOY) parr was tested in the River Dalåa from mid-May to late-June 1999. Hypo-osmoregulatory capacity, measured as plasma osmolality and chloride, was assessed after seawater (SW) challenge tests (168 h, salinity = 35). All S. salar exhibited sufficient hypo-osmoregulatory capacity at the initiation of downstream migration in mid-May. Migrating S. salar smolts caught in mid-May and retained in fresh water displayed no signs of de-smoltification as they maintained hypo-osmoregulatory capacity through June. This indicates a physiological smolt window that lasts a minimum of 6 weeks (330 degree days; D°) for hatchery-produced S. salar smolts stocked as YOY parr. Based on the observed river migration speeds, it can be assumed that the S. salar smolts entered SW 2-4 weeks after initiation of migration in the upper parts of the River Dalåa. Hence, based on smolt migration and SW tolerance, it is suggested that stocking of YOY parr is a viable enhancement strategy in the River Dalåa.