Feminizing endocrine disrupting compounds (EDCs) affect the growth and development of teleost fishes. The major regulator of growth performance, the growth hormone (Gh)/insulin-like growth-factor (Igf) system, is sensitive to estrogenic compounds and mediates certain physiological and potentially behavioral consequences of EDC exposure. Igf binding proteins (Igfbps) are key modulators of Igf activity, but their alteration by EDCs has not been examined. We investigated two life-stages (fry and smolts) of Atlantic salmon (Salmo salar), and characterized how the Gh/Igf/Igfbp system responded to waterborne 17α-ethinylestradiol (EE2), 17β-estradiol (E2) and 4-nonylphenol (NP). Fry exposed to EE2 and NP for 21 days had increased hepatic vitellogenin (vtg) mRNA levels while hepatic estrogen receptor α (erα), gh receptor (ghr), igf1 and igf2 mRNA levels were decreased. NP-exposed fry had reduced body mass and total length compared to controls. EE2 and NP reduced hepatic igfbp1b1, -2a, -2b1, -4, -5b2 and -6b1, and stimulated igfbp5a. In smolts, hepatic vtg mRNA levels were induced following 4-day exposures to all three EDCs, while erα only responded to EE2 and E2. EDC exposures did not affect body mass or fork length; however, EE2 diminished plasma Gh and Igf1 levels in parallel with reductions in hepatic ghr and igf1. In smolts, EE2 and E2 diminished hepatic igfbp1b1, -4 and -6b1, and stimulated igfbp5a. There were no signs of compromised ionoregulation in smolts, as indicated by unchanged branchial ion pump/transporter mRNA levels. We conclude that hepatic igfbps respond (directly and/or indirectly) to environmental estrogens during two key life-stages of Atlantic salmon, and thus may modulate the growth and development of exposed individuals.

Growth performance of teleost fishes, including salmonids, is principally controlled by the growth hormone (Gh)/insulin‐like growth‐factor (Igf) axis. Igfs interact with cognate binding proteins, termed Igf binding proteins (Igfbps), which modulate the biological activities of Igfs. While Atlantic salmon (Salmo salar) are known to express an expansive set of igfbp transcripts in several key metabolic tissues, the actual regulators of igfbp expression have not been identified. Given the well‐documented consequences of mounting a glucocorticoid stress response on growth, we hypothesized that cortisol inhibits anabolic processes by directing the dynamic expression of igfbps. We therefore examined changes in the activities of the Gh/Igf/Igfbp network of salmon parr implanted with cortisol (0, 10 and 40 μg/g body weight) and sampled after 3 or 14 days. We observed reductions in body mass and fork length following 3‐ and 14‐day treatments with cortisol at 40 μg/g. A dose dependent reduction in growth rate occurred by 14 days of treatment with cortisol, whereas hepatosomatic index was not affected. We will report igfbp transcript levels in liver, skeletal muscle and gill with an aim to resolve how activities within the Igfbp network are modulated by cortisol in juvenile Atlantic salmon.Support or Funding InformationSupported by Start‐Up funds to J.P.B.

Specific growth hormone (GH)-binding protein (Ghbp) was purified from Atlantic salmon Salmo salar and rainbow trout Oncorhynchus mykiss plasma with immunoprecipitation and characterized in cross-linking studies using autoradiography and western blots. The size of the Ghbp was estimated to be c. 53 kDa. A radioimmunoassay was established to measure Ghbp in salmonids, using antibodies specific against the extracellular segment of the S. salar growth hormone receptor 1 (grh1; GenBank AY462105). Plasma Ghbp levels were measured in S. salar smolts in fresh water and after transfer to seawater (SW; experiments 1 and 2), and in post-smolts kept at different salinities (0, 12, 22 and 34) for 3 months (experiment 3). A transient increase in plasma Ghbp, which lasted for 1 month or less, was noted in smolts after transfer to SW. Concomitantly, plasma GH and gill Na(+) -K(+) -ATPase activity increased during smoltification (in experiment 2). No difference in plasma Ghbp was evident between post-smolts kept at different salinities, although the fish kept at salinity 34 had higher plasma GH than the group kept at salinity 22 and higher hepatic ghr1 expression than post-smolts kept at salinity 12. This suggests that plasma Ghbp regulation may respond to salinity changes in the short term. The lack of correlation between Ghbp, plasma GH and hepatic ghr1 expression in the long-term post-smolt experiment indicates that Ghbp levels may be regulated independently of other components of the endocrine GH system in salmonids.

Insulin was purified from pancreatic extracts of two elasmobranch species belonging to different families in the order Carcharhiniformes, the European spotted dogfish, Scyliorhinus canicula (Scyliorhinidae), and the hammerhead shark, Sphyrna lewini (Carcharhinidae). The amino acid sequence of dogfish insulin was established as A-chain GIVDHCCRNT(10)CSLYDLEGYC(20)NQ and B-chain LPSQHLCGSH(10)LVETLYFVCG(20)QKGFYYVPKV(30). The primary structure of hammerhead shark insulin was similar to that of dogfish insulin with only 2 amino acid substitutions at A8 (R --> H) and B30 (V --> I). The elasmobranch insulins were markedly different from human insulin (17 amino acid substitutions) but all the residues in human insulin that are believed to be important in determining the receptor binding conformation (B6, B8, B11, B13, B23, B24, B25, A2, A3, and A19) have been conserved in the elasmobranch insulins with the exception of the conservative substitution Phe --> Tyr at B25. Consistent with this, dogfish and human insulin showed almost identical binding affinity to the recombinant solubilized human insulin receptor (K(D) values of 14.0 and 18.6 pM, respectively; relative potency 133%). Previous studies have shown that bovine insulin produces severe and sustained hypoglycemia in elasmobranchs but the effect is of slow onset. Bolus arterial injections of dogfish insulin (10 nmol x kg(-1)) into unanesthetized, fasting dogfish (n = 9) produced no changes in blood glucose, 3-hydroxybutyrate, and acetoacetate concentrations over a 4-h period. In a second series of experiments (n = 7), dogfish insulin (10 nmol x kg(-1)) produced a significant (P < 0.05) fall in blood glucose after 12 h that persisted for at least 48 h, but no change in ketone body concentrations. The data indicate that the metabolic actions of an endogenous elasmobranch insulin in an elasmobranch are similar to those previously described for mammalian insulin.

The main objectives of this study were to obtain the amino acid sequence of Atlantic halibut (Hippoglossus hippoglossus) growth hormone (hhGH) and compare it with other teleost species, to establish a radioimmunoassay to assess plasma hhGH levels and thus to gain information about possible biological functions and regulation by photoperiod. The hhGH gene was cloned and its amino acid sequence deduced from the cDNA. The mature hhGH protein consists of 186 amino acids. Comparison with other flatfish species as well as a species from a different order, the pufferfish, reveals that the sequence similarities of the mature hhGH with that of the barfin flounder, the Japanese flounder, the sole and the pufferfish are 99.5, 81.7, 74.2, and 65.2%, respectively. The sequence similarities appear to correctly reflect the gross phylogenetic relationships among these teleost species. A specific GH-RIA was developed for measurements of Atlantic halibut GH levels. Assessment of plasma GH levels in adult halibut revealed large gender differences, with GH levels frequently being an order of magnitude higher in males than females. The mean (+/-SEM) plasma GH for males kept on normal annual photoperiod were 25.2+/-6.11 ngml(-1) and for females were 5.14+/-1.94 ngml(-1). It appears likely that plasma growth hormone levels in Atlantic halibut can be inversely correlated to growth and metabolism. Shifting of the annual photoperiod cycles demonstrated that photoperiod in not a regulator of plasma GH levels in the Atlantic halibut, but further research is needed to assess whether GH plays a role in the reproduction of this marine teleost species.

The thyroid hormones (THs), thyroxine (T(4)) and triiodothyronine (T(3)) are products of the thyroid gland in all vertebrates. Their role in early development and metamorphosis is well established in mammals and amphibians, respectively, and recently several studies in fish have highlighted the importance of THs during flatfish metamorphosis. THs are present in high quantities in fish eggs and are presumably of maternal origin. During embryogenesis the concentration of T(4) and T(3) in the eggs decrease until endogenous production starts. Thyroid hormone receptors (TR) have been isolated from several teleosts and in common with tetrapods two receptor isoforms have been identified, TR alpha and TR beta. Both the receptors are expressed in early embryos and larvae of the Japanese flounder (Paralichthys olivaceus), zebrafish (Danio rerio) and seabream (Sparus aurata) although a different temporal pattern is apparent. The role of THs and TRs in fish embryogenesis, larval development and during metamorphosis will be discussed.

This study was carried out to test the effects of acute stress on the primary and the secondary antibody responses in Atlantic salmon (Salmo salar L.). Fish received a primary immunization with sheep red blood cells (SRBC) and received a second immunization 11 weeks later. At priming, a group of fish was stressed by water level reduction for 30 min (moderate stress), and another group was stressed severely by applying the stressor twice, with an interval of 6 h, 3 days in a row (severe stress). At the time of the booster, another two groups of previously unstressed, primed fish were treated likewise. Unstressed fish mounted a primary antibody response to SRBC and a secondary response, which was improved compared with the primary response., The primary antibody response against SRBC was impaired by severe, but not moderate, acute stress at priming, whereas the secondary antibody response was unaffected. Both moderate and severe stress at the time of the booster was immunosuppressive in fish that had been primed under non-stress conditions. It is concluded that cortisol release as a result of stress exposure is most immunosuppressive at the time of the second immunization.

White muscle high-energy phosphate levels showed that the bleeding procedure applied during slaughter of Atlantic salmon (Salmo salar) did not overshadow the total pre-slaughter handling stress. The energy status of the muscle seemed to increase as a result of bleeding and the bleeding per se did not have adverse effects on flesh freshness resulting from excessive handling stress. The time for onset of rigor mortis and rigor strength were clearly related to the initial stress level and the rate of ATP depletion. The onset of rigor in stressed fish occurred about one day earlier compared with rested fish and the stressed fish also became stiffer.