Characterization of growth hormone binding sites in the goldfish,Carassius auratus: effects of hypophysectomy and hormone injection

A recombinant carp growth hormone (rcGH) was used to develop for a GH radioreceptor binding assay in the goldfish (Carassius auratus). Specific binding of¹²⁵I-rcGH to goldfish liver membranes was a pH, time, temperature, and membrane protein dependent process. Scatchard and LIGAND analysis indicated a single class of high affinity and low capacity binding site, with an association constant (Ka) of 1.9×10¹⁰ M^–1 and a maximum binding capacity (B_max) of 9 fmol mg^–1 protein. Liver tissue displayed the highest¹²⁵I-rcGH binding of all the tissues examined. Displacement of¹²⁵I-rcGH with various unlabeled teleost and mammalian GHs and prolactins revealed that the goldfish hepatic binding site was highly specific for teleost GH. Intraperitoneal administration of 0.1, 1.0, and 10 g rcGH g^–1 body weight to hypophysectomized goldfish resulted in a 27, 52, and 68% decrease in total binding sites, respectively. Injection of a high dose of rat prolactin (rPRL) (5 g rPRL g^–1 body weight) also resulted in a 32% decrease in total binding sites. These results suggest that endogenous GH may have a role in the regulation of its own receptors in the goldfish.     

Quantitation of inert feed ingestion in larval silver sea bream (Sparus sarba) using auto-fluorescence of alginate-based microparticulate diets

The ingestion of an inert feed as a sole food source was investigated in larval silver sea bream (Sparus sarba) fed an alginate-based microparticulate diet. Using the auto-fluorescent properties of pigments associated with the alginate base, ingestion and gut content were investigated over a 6 h experimental period in fed and unfed larvae. By extracting and measuring chlorophyll a (Chl a) and phaeopigment content of feeding larval fish and relating this to standardized Chl a and phaeopigment content of the diet, relative to diet weight, it was determined that individual fed 7-day old larvae had a maximum gut content of 1.05±0.09 g diet while 14-day old fed fish had a maximum gut content of 3.17±0.90 g diet. On average, the gut content of 14-day old fish was 2.89 times greater than the gut content of 7-day old fish. The dry weight of larval sea bream increased from 43±4.2 g at day 7 to 134.3±20.4 g at day 14 indicating that growth of fish fed this inert feed was substantial. Gut pigment dynamics suggested that Chl a was degraded to phaeopigments by 7-day but not 14-day old larvae and the individual gut dietary content varied considerably in 14-day old fish. The maximum Chl a and phaeopigment content in larval sea bream was 0.4 ng ind^–1 and 0.55 ng ind^–1 for 7-day old fish and 1.54 ng ind^–1 and 2.81 ng ind^–1 for 14-day old fish respectively. The present method may potentially allow simple and direct assessment of larval fish feed ingestion in both an experimental and commercial setting.     

Insulin-like growth factor I (IGF-I) mRNA expression in coho salmon,Oncorhynchus kisutch: Tissue distribution and effects of growth hormone/prolactin family proteins

To examine the hormonal and nutritional regulation of insulin-like growth factor I (IGF-I) mRNA expression, a sequence-specific solution hybridization/RNase protection assay for coho salmon IGF-I mRNA was developed. This assay is both rapid and sensitive and has low inter- (less than 15%) and intra-assay variations (less than 5%). Using this assay, the tissue distribution of IGF-I mRNA and effects of growth hormone (GH), prolactin (PRL) and somatolactin (SL) on hepatic IGF-I mRNA expression in coho salmon were examined in vivo. Liver had the highest IGF-I mRNA level of 16 pg/μg DNA. Significant amounts of IGF-I mRNA were also found in all other tissues examined (intestine 4.1, kidney 3.8, gill arch 2.4, brain 2.4, ovary 2.3, muscle 2.1, spleen 1.7 and fat 1.1 pg/μg DNA). Injection of coho salmon GH at doses of 0.1 and 1 μg/g body weight significantly increased the hepatic IGF-I mRNA levels in a dose-dependent manner. Injection of coho salmon SL, a recently discovered member of the GH/PRL family, stimulated the IGF-I mRNA expression at the higher dose (1 μg/g), whereas coho salmon PRL had no effect at either dose. Concentration-dependent stimulation by coho salmon GH was also obtained in vitro in primary culture of salmon hepatocytes in concentrations ranging from 0.01 to 1 μg/ml. These results indicate that IGF-I mRNA expression occurs in a variety of tissues in coho salmon, and that at least the hepatic expression is under the regulation of GH and possibly other hormones. The sequence-specific assay established in the present study can be used for accurate quantitation of IGF-I mRNA in salmonid species, and can contribute to a better understanding of the physiology of IGF-I in salmonids.

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Résumé Afin d'étudier les régulations homronales et nutritionnelles de l'expression des ARNm de l'IGF-I (insulin-like growth factor I), un dosage spécifique par hybridation en solution des ARNm d'IGF-I de saumon coho et protégé des RNases, a été développé. Ce dosage, à la fois rapide et sensible, présente un faible coefficient de variation inter- (< 15%) et intra- (< 5%) dosage. L'étude de la distribution tissulaire des ARNm de l'IGF-I et des effets de l'hormone de croissance (GH), de la prolactine (Prl) et de la somatolactine (SI) sur l'expression hépatique des ARNm de l'IGF-I, a été entreprise in vivo chez le saumon coho en utilisant ce dosage. Le foie présente les plus grandes quantités d'ARNm d'IGF-I (16 pg/μg d'ADN). Des quantités significatives d'ARNm d'IGF-I ont été également détectées dans tous les autres tissus étudiés (intestin 4,1; rein 3,8; branchie 2,4; ovaire 2,3; muscle 2,1; rate 1,7 et graisse 1,1 pg/μg d'ADN). L'injection à des saumons coho, de GH à des doses de 0,1 et 1 μg/g de poids vif, augmente significativement et de manière dose dépendante les niveaux hépatiques d'ARNm d'IGF-I. L'injection de SI de saumon coho, un membre récemment découvert de la famille GH/Prl, stimule avec la plus haute dose utilisée, l'expression des ARNm d'IGF-I alors que la Prl n'a aucun effet. La GH augmente de manière dose dépendante (0,01–1 μg/ml) l'expression in vitro des ARNm d'IGF-I par des ARNm d'IGF-I par des hépatocytes de saumon coho en culture. Ces résultats indiquent que, chez le saumon coho, l'expression des ARNm d'IGF-I est présente dans le nombreaux tissus et que, l'expression hépatique est, au moins en partie, régulée par la GH et peut-être par d'autres hormones. Le dosage par séquence spécifique mise au point dans le présent travail, peut-être utilisé pour la quantification précise des ARNm, d'IGF-I de salmonidés et devrait permettre une meilleure connaissance de la physiologie de L'IGF-I chez les salmonidés.

     

In vivo and in vitro effect of recombinant salmon growth hormone treatment on IGF-I and IGFBPs in yellowtail <Emphasis Type="Italic">Seriola quinqueradiata</Emphasis>

The role of growth hormone (GH) in regulating hepatic mRNA expression of insulin-like growth factor-I (IGF-I) and IGF binding proteins (IGFBPs) in yellowtail Seriola quinqueradiata was examined using in vivo and in vitro assays. Yellowtail hepatic IGF-I, IGFBP-1, IGFBP-2, IGFBP-3, and IGFBP-5 mRNAs were measured by real-time quantitative RT-PCR. Intraperitoneal injection of recombinant GH of chum salmon Oncorhynchus keta (rsGH) at a dose of 1 μg/g body weight resulted in a significant increases in hepatic IGF-I, IGFBP-3, and IGFBP-5 mRNA levels, whereas significant reductions in hepatic IGFBP-1 and IGFBP-2 mRNA levels were observed. For in vitro assays, liver slices were incubated with rsGH at different concentrations (doses: 0, 1, 10, 100, 500, and 1,000 ng/ml). Liver slices incubated with 100 ng/ml rsGH elicited a significant increase in IGF-I mRNA level. Similarly, a slight increase in IGFBP-3 and IGFBP-5 mRNA levels were also observed in liver explants incubated with rsGH. In contrast, a significant decline in IGFBP-1 mRNA levels was observed in liver slices incubated with 1,000 ng/ml rsGH. A slight decline in the level of IGFBP-2 mRNA was noted in liver explants with rsGH treatment. This study demonstrates the modulating effect of GH on the IGF system.     

Dopamine inhibits gonadotropin secretion in the Chinese loach (Paramisgurnus dabryanus)

The effects of dopamine on gonadotropin (GtH) secretion in sexually mature Chinese loach were investigated. Spontaneous secretion of GtH was inhibited within 1 h following an intramuscular injection of dopamine (100 g/g body wt). Similarly, dopamine (50 and 100 g/g body wt) caused a significant reduction in serum GtH in fish with elevated GtH levels as a result of pretreatment with gonadotropin-releasing hormone (GnRH) analogs either alone or in combination with the dopamine receptor antagonist domperidone. In summary, the present study provides direct evidence that dopamine functions as a gonadotropin-release inhibitory factor in the Chinese loach by blocking spontaneous and GnRH-stimulated GtH release.     

Angiotensins stimulate in vitro ovulation and contraction of brook trout (Salvelinus fontinalis) follicles

The effects of salmon angiotensin I (sAI) and human angiotensin II (hAII) on in vitro ovulation of preovulatory (preOV) brook trout (Salvelinus fontinalis) follicles were investigated. Both angiotensins increased levels of ovulation above that in controls after 12 hours of incubation. The increase was statistically significant in incubates with greater than 1 M hAII. The effects of the angiotensins on follicle contraction were also studied indirectly by measuring the decrease in weight of punctured follicles taken prior to germinal vesicle breakdown. At 1 M, both angiotensins significantly decreased the weight of punctured follicles after 16 hours of incubation. The angiotensin-stimulated decrease in weight was not blocked by indomethacin (10 g/ml), indicating that follicle contraction was not prostaglandin-dependent. The data indicate that angiotensins might be directly involved in brook trout ovulation and the stimulatory effects of angiotensins on ovulation may be attributed to their effects on follicle contraction.This work was supported by NIH grant #HD25924-02.     

Disparate effects of gonadal steroid hormones on plasma and liver mRNA levels of insulin-like growth factor-I and vitellogenin in the tilapia, Oreochromis mossambicus

The tilapia, Oreochromis mossambicus, exhibits a sexually dimorphic pattern of growth, males growing larger than females. We examined the effects of E₂ and DHT on the GH/IGF-I axis and on VTG production in the tilapia. Sexually mature tilapia were injected with 5 μg g body weight of E₂ (males) or DHT (females) every 5 days for a total of 3 injections. Female tilapia had significantly higher plasma GH levels than males. However, plasma and liver mRNA levels of IGF-I were significantly lower in females than in males, whereas VTG levels in both the plasma and liver mRNA were significantly higher in females than in males. Although significant amounts of VTG were detected in control males (8 ± 0.3 μg ml), the levels in control females (3000 ± 500 μg ml) were about 400 times higher than in males. Males treated with E₂ exhibited a female-like GH/IGF-I profile. That is, they had significantly elevated levels of plasma GH with lower plasma IGF-I and liver IGF-I mRNA levels. Estradiol treatment significantly elevated both plasma and liver mRNA VTG levels. Dihydrotestosterone treatment in females induced a male-like GH/IGF-I profile: plasma GH levels were significantly reduced, whereas plasma and liver IGF-I mRNA levels were significantly elevated. Both plasma and liver mRNA levels of VTG were not altered by DHT treatment. Pituitary GH mRNA levels were similar in all treatment groups. These results clearly indicate that estrogens and androgens feminize and masculinize the GH/IGF-I axis, respectively. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of triiodothyronine and propylthiouracil on thyroid function and smoltification of coho salmon (Oncorhynchus kisutch)

Yearling coho salmon (Oncorhynchus kisutch) were fed diets containing triiodothyronine (T₃; 4 or 12gmg), propylthiouracil (PTU; 1.5 or 6.0 mg/g), or both T₃ (12g/g) and PTU (1.5 mg/g) from January 10 to May 29. Plasma T₄ and T₃, concentrations were maintained within normal physiological limits in all groups of treated fish. Increases in plasma thyroxine (T₄) occurred in late April in groups receiving the high dose of T₃ or PTU, or the combination of T₃, and PTU. Peaks of plasma T₄ occurred in May in the other groups. Thyroid follicle epithelial cell height was increased in the groups in the following order: highest; PTU (6.0 mg/g), PTU (1.5 mg/g), PTU + T₃, control, T₃ (4gmg/g), T₃ (12g/g); lowest. In March specific binding of T₃ by liver nuclei was not affected by treatment with T₃ (12g/g) but was decreased 30% by treatment with PTU (6.0g/g). Body growth, food conversion efficiency, and pigmentary changes were increased by T₃ (12g/g), decreased dose-dependently by PTU, and unaffected by the other dietary treatments. All treatments altered body proximate composition. Food consumption and the ability to osmoregulate in seawater were decreased in fish fed 6.0 mg/g PTU, but were unaffected by the other dietary treatments. These results suggest that during smoltification: 1) Thyroid hormones may be involved in control of thyroidal function and body growth and pigmentary changes, 2) Hypo-osmoregulatory competence is not further stimulated by exogenous T₃ and 3) PTU does not block T₄ synthesis but may block T₃, action by interfering with nuclear T₃ binding.     

Involvement of gonadal steroids in final oocyte maturation of white perch (Morone americana) and white bass (M. chrysops): in vivo and in vitro studies

Plasma estradiol-17 (E₂), testosterone (T), 17,20-dihydroxy-4-pregnen-3-one (DHP) and 17,20,21-tri-hydroxy-4-pregnen-3-one (20-S) levels were measured by radioimmunoassay (RIA) in white perch (Morone americana) and white bass (M. chrysops) that were induced to undergo final oocyte maturation (FOM) with human chorionic gonadotropin (hCG). Plasma DHP levels increased in females of both species in association with oocyte germinal vesicle migration (GVM) and germinal vesicle breakdown (GVBD) and decreased thereafter. Plasma 20-S levels also increased with oocyte GVM in white bass, but were several-fold lower than DHP levels. Circulating E₂ and T levels were greatest during GVM and GVBD in both species and decreased to low levels during oocyte hydration and ovulation. Follicles from white perch and white bass which received a priming injection of hCG in vivo, produced both DHP and 20-S in vitro after exposure to hCG and their oocytes underwent GVBD. Ovarian incubates from unprimed fish of either species produced only E₂ and T and their oocytes did not complete GVBD. Oocytes from unprimed bass, but not perch, matured when follicles were exposed to hCG in vitro. Both trilostane and cycloheximide blocked in vitro production of DHP and 20-S and oocyte GVBD by white perch follices. DHP and 20-S were equipotent inducers of FOM in the GVBD bioassay. None of several other structurally-related steroids tested were effective within a physiological range of concentrations. These results indicate a role for DHP and 20-S in the control of FOM in white perch and white bass.     

17β-Estradiol, but not testosterone stimulates gonadotropin II concentrations in the protandrous black porgy, Acanthopagrus schlegeli Bleeker

The objective of the present study was to investigate the in vivo effects of different doses of 17-estradiol (E₂) and testosterone (T) on the levels of plasma and pituitary gonadotropin II (GTH II) in 2-year-old black porgy, Acanthopagrus schlegeli, during the spawning season. Male fish were distributed among 7 groups (n = 49), control, E₂or T (with 3 doses, 2.4 ng, 72 ng and 2.2 g g^–1 body weight). Fish were injected with the respective vehicle or different doses of E₂ or T on days 1 and 14. Plasma E₂ levels were significantly increased in the 72 ng E₂ group on days 8 and 14. Plasma vitellogenin levels were significantly higher in the 72 ng E₂ group on days 14 and 20, and 2.2 g E₂ group on days 8, 14 and 20 than those in the control group. Plasma GTH II concentrations were significantly higher in the 2.2 g E₂ group than in the control and other E₂groups on days 8, 14 and 20. Pituitary GTH II contents was significantly higher in the 7.2 ng E₂ group compared to the control and other E₂groups on day 20. Plasma GTH II concentrations were similar in the control and all the T groups on days 8, 14 and 20. None of the doses of T treatment stimulated pituitary GTH II content on day 20, although plasma vitellogenin levels were elevated. It is concluded that GTH II synthesis and secretion in black porgy is stimulated by an estrogen-specific effect.     

Catecholestrogens inhibit dopamine methylation in the gonadotrops of the African catfish,Clarias gariepinus

Isolated gonadotrops of the African catfish,Clarias gariepinus, were incubated with dopamine (DA) and/or catecholestrone and the activity of the enzyme catechol-O-methyltransferase (COMT) was determined by measuring the methylated products. From the apparent Km values for DA and catecholestrone of 0.4–1.3 M and 17.9–25.2 M respectively, it was concluded that catecholestrone is a better substrate for the enzyme COMT, compared to DA. Moreover, the methylation of DA is inhibited by comparatively low concentrations of catecholestrone.     

Antisomatostatin-induced growth acceleration in chinook salmon (Oncorhynchus tshawytscha)

Since somatostatin (SRIF) inhibits the release of growth hormone (GH), its immunoneutralization may provide an alternative to GH therapy as a means of enhancing somatic growth in fish. The present study examined the feasibility of accelerating growth in juvenile chinook salmon by means of antiSRIF administration. Yearling salmon of Nicola River stock (BC, Canada) were injected intraperitoneally every 5 days, for a total of 40 days, with either SRIF (1 μg g-1 body wt.), antiSRIF (SOMA-10, 1 μg g⁻¹), recombinant bovine GH (rbGH, 2.5 μg g⁻¹), recombinant porcine GH (rpGH, 2.5 μg g⁻¹) or saline (controls). No significant differences were observed in length, weight or final condition factor (k) between the SRIF-treated and control fish over the experimental period. However, the fish treated with the antiSRIF were significantly (p ≤ 0.05) longer and heavier than the control salmon after 25 and 30 days respectively. Furthermore, antiSRIF treatment caused a lowering in k when compared to the control salmon. Fish injected with rbGH or rpGH were significantly longer and heavier than all other groups (p ≤ 0.05), after only 5 days. GH treated groups also returned higher k when compared against all other treatments (p ≤ 0.05). No differences were observed in growth between the two rGH treatments over the experimental period.     

Influence of cortisol, growth hormone, insulin-like growth factor I and 3,3′,5-triiodo-l-thyronine on hypoosmoregulatory ability in the euryhaline teleost Fundulus heteroclitus

The capacity of cortisol, ovine growth hormone (oGH), recombinant bovine insulin-like growth factor I (rbIGF-I) and 3,3,5-triiodo-l-thyronine (T₃) to increase hypoosmoregulatory capacity in the euryhaline teleost Fundulus heteroclitus was examined. Fish acclimated to brackish water (BW, 10 ppt salinity) were injected with a single dose of hormone suspended in oil and transferred to seawater (SW, 35 ppt salinity) 10 days post-injection. Fish were sampled 24 h after transfer and plasma osmolality and gill Na⁺, K⁺-ATPase activity were examined. Transfer from BW to SW induced significantly increased plasma osmolality but not gill Na⁺, K⁺-ATPase activity. Cortisol (50 g g^–1 body weight) improved the ability to maintain plasma osmolality and to increase gill Na⁺, K⁺-ATPase activity. oGH (5 g g^–1 body weight) also increased hypoosmoregulatory ability and gill Na⁺, K⁺-ATPase activity. A cooperation between oGH and cortisol was observed in increasing hypoosmoregulatory ability but not in increasing gill Na⁺, K⁺-ATPase activity. rbIGF-I (0.5 g g^–1 body weight) alone was without effect in increasing salinity tolerance or gill Na⁺, K⁺-ATPase activity. rbIGF-I and oGH showed a positive interaction in increasing salinity tolerance, but not gill Na⁺, K⁺-ATPase activity. Treatment with T₃ (5 g g^–1 body weight) alone did not increase salinity tolerance or gill Na⁺, K⁺-ATPase activity, and there was no consistent significant interaction between cortisol and T₃ or between GH and T₃. The results confirm the classical role of cortisol as a seawater-adapting hormone and indicate an interaction between cortisol and the GH/IGF-I axis during seawater acclimation of Fundulus heteroclitus.     

The effects of gonadal development and sex steroids on growth hormone secretion in the male tilapia hybrid (Oreochromis niloticus × O. aureus)

Profiles of plasma growth hormone (GH) in male tilapia hybrid (Oreochromis niloticus x O. aureus) were measured and compared at different times of the year. The profiles did not appear to be repetitive, however, differences in their nature were observed at the different seasons; the most erratic profiles were seen in the height of the reproductive season (July), while the peaks were more subdued in the spring and disappeared in the autumn. Peaks in male fish were more prominent than in the females when measured in July. Perifused pituitary fragments from fish with a high GSI responded to salmon gonadotropin-releasing hormone (sGnRH) analog (10 nM-1 M), while those from fish with a low GSI barely responded to even the highest dose. Exposure of perifused pituitary fragments from sexually-regressed fish to carp growth hormone-releasing hormone (cGHRH; 0.1 M) or sGnRH (I M) stimulated GH release only after injection of the fish with methyl testosterone (MT; 3 injections of 0.4 mg kg ¹). The same MT pretreatment did not alter the response to dopamine (DA; 1 or 10 M). GH pituitary content in MT-treated fish was lower than in control fish, which may be explained by the higher circulating GH levels in these fish, but does not account for the increased response to the releasing hormones. Castration abolished the response of cultured pituitary cells to sGnRH (I fM-100 nM) without altering either their basal rate of secretion or circulating GH levels. Addition of steroids to the culture medium (MT or estradiol at 10 nM for 2 days) enabled a GH response to sGnRH stimulation in cells from sexually regressed fish. Pituitary cells which had not been exposed to steroids failed to respond to sGnRH, although their response to forskolin or TPA was similar to that of steroid-exposed cells. It would appear, therefore, that at least one of the effects of the sex steroids on the response to GnRH is exerted proximally to the formation of cAMP, or PKC, presumably at the level of the receptor. An increase in the number of receptors to the GH-releasing hormones, following steroid exposure, would explain also the changing nature of the GH secretory profile in different stages of the reproductive season.     

Evidences for involvement of growth hormone and insulin-like growth factor in ovarian development of starry flounder (<Emphasis Type="Italic">Platichthys stellatus</Emphasis>)

Although gonadotrophins are major regulators of ovarian function in teleosts and other vertebrates, accumulating evidence indicates that the growth hormone (GH)-insulin-like growth factor (IGF) axis also plays an important role in fish reproduction. As a first step to understand the physiological role of the GH-IGF system in the ovarian development of starry flounder (Platichthys stellatus), the expression profiles of GH and IGF messenger RNAs (mRNAs) and plasma GH, IGF-I, estradiol-17β (E2), and testosterone (T) levels during the ovarian development were investigated. The developmental stages of ovaries were divided into five stages (II, III, IV, V, and VI) by histological analysis. The hepatosomatic index (HSI) and gonadosomatic index (GSI) values increased and peaked at stage IV and stage V, respectively, and then declined at stage VI. Pituitary GH mRNA levels decreased sharply at stage III and raised to top level at stage VI. The hepatic IGF-I mRNA levels ascended to maximum value at stage V and then declined significantly at stage VI. However, the hepatic IGF-II mRNA levels remained stable and increased significantly at stage VI. In contrast, the ovarian IGF-I mRNA levels increased gradually and peaked at stage VI. The ovarian IGF-II mRNA levels were initially stable and increased significantly at stage V until the top level at stage VI. Consistent with the pituitary GH mRNA levels, plasma GH levels reduced sharply at stage III and remained depressed until stage V and then raised remarkably at stage VI. Plasma IGF-I level peaked at stage V and then declined to initial level. Plasma E2 level peaked at stage IV and then dramatically descended to the basal level. Plasma T level peaked at stage V and then declined significantly back to the basal level. Based on statistical analysis, significant positive correlations between hepatic IGF-I mRNA and GSI, ovarian IGF-II mRNA and hepatic IGF-II mRNA, ovarian IGF-I mRNA and ovarian IGF-II mRNA, and plasma IGF-I and plasma T were observed, respectively. These results suggest that the GH-IGF system may be involved in the ovarian development of starry flounder; GH and IGFs appear to play distinct roles in the regulation of the ovarian development in paracrine/autocrine manners. These findings extend our knowledge of the roles of the GH-IGF axis on reproduction regulation in fish.     

Effect of d-Lys6 salmon sGnRH alone and in combination with domperidone on the spawning of common carp during the late spawning season

The effect of [d-Lys6] salmon gonadotropin-releasing hormone alone (sGnRH) and in combination with domperidone, a dopamine antagonist, on the spawning of common carp, Cyprinus carpio, was investigated during the late spawning season (when mature fish stop spawning under natural conditions when males and females are put together). Fish were induced to spawn by [d-Lys6]-sGnRH salmon gonadotropin-releasing hormone analogue (sGnRH-X) only when injected in combination with domperidone at doses of 10 g kg-1 and 20 mg kg-1 body weight respectively. [d-Lys6]-sGnRH-X alone did not induce spawning at a dose as high as 100 g kg-1 body weight. Fish injected with domperidone alone did not spawn either. These results demonstrate that [d-Lys6]-sGnRH-X in combination with a dopamine antagonist is effective in inducing spawning during the late spawning season.     

Seasonal changes in circulating growth hormone (GH), hepatic GH-binding and plasma insulin-like growth factor-I immunoreactivity in a marine fish,gilthead sea bream,Sparus aurata

We have studied the seasonal relationship between growth and circulating growth hormone (GH), hepatic GH-binding and plasma insulin-like growth factor-I immunoreactivity in gilthead sea bream,Sparus aurata. The seasonal increase in plasma GH levels preceded by several weeks the summer increase in growth rates. In contrast, a marked increase in hepatic GH-binding with a high degree of endogenous GH occupancy was found during the period of maximum growth which suggests an enhanced sensitivity of liver to GH action. Thus, circulating levels of immunoreactive IGF-I, probably derived from the liver in response to GH action, were positively correlated with growth throughout the experimental period although a consistent relationship between growth and circulating GH was not found. In spite of this, we consider that, in gilthead sea bream, as in several other teleosts, the availability of endogenous GH can limit growth. Thus, under environmental conditions of suboptimal growth, a single intraperitoneal injection of recombinant rainbow trout GH (rtGH) induced over the dose range tested (0.75, 1.5, 3 μg g BW⁻¹) an increase in plasma IGF-I-like immunoreactivity comparable to that seen during the period of maximum growth.     

The presence of uric acid, an antioxidantive substance, in fish seminal plasma

High concentrations of uric acid in seminal plasma of a range of teleost fish species are reported for the first time. Concentrations of urate amounted to 223.4; 121.9–130.0, 355.9, 735.6, 124.0, 192.7 and 148.0 M for rainbow trout (Oncorhynchus mykiss), yellow perch (Perca flavescens), muskellunge (Esox masquinongy), Northern pike (Esox lucius), carp koi (Cyprinus carpio), bream (Abramis brama), and tench (Tinca Tinca), respectively. Uric acid is an important anti-oxidant, therefore, it has a potential role in protection of fish spermatozoa against oxidative damage.     

Effects of purified microcystin-LR and cell extracts of Microcystis strains PCC 7813 and CYA 43 on cardiac function in brown trout (Salmo trutta) alevins

Sub-lethal cardiac responses of brown trout alevins (Salmo trutta L.) were determined in response to aqueous extracts of the cyanobacterium Microcystis strains PCC 7813 (microcystins detectable by HPLC) and CYA 43 (no microcystins detectable by HPLC) and to the purified cyanobacterial hepatotoxin, microcystin-LR (MC-LR) at concentrations of 5, 50 and 500 g microcystin-LR equivalents l^–1. Responses were determined using a flow chamber and video camera attached to a low power microscope. Heart rate in brown trout alevins was acutely sensitive to cyanobacterial extracts and significant increases were observed within 15–60 sec of exposure to aqueous extracts, although no change was observed on exposure to purified MC-LR. Stroke volume increased in all treatments at 50 and 500 g MC-LR equivalents l^–1, which may, at least in part, have been due to vasodilation of the yolk-sac blood vessels. Cardiac output increased significantly at all three concentrations of cyanobacterial cell extracts but not at the lowest concentration of MC-LR, although the rate increased at levels at/or above 50 g l^–1. Increased heart rate, stroke volume and cardiac output occurred at environmentally relevant microcystin concentrations of Microcystis PCC 7813 and CYA 43 aqueous extracts.