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1.
The duality of teleost gonadotropins   总被引：5，自引：0，他引：5  
The duality of salmon gonadotropins has been proved by biochemical, biological, and immunological characterization of two chemically distinc gonadotropins. GTH I and GTH II were equipotent in stimulating estradiol production, whereas GTH II appears to be more potent in stimulating maturational steroid synthesis. The ratio of plasma levels and pituitary contents of GTHs and the secretory control by a GnRH suggest that GTH I is the predominant GTH during vitellogenesis and early stages of spermatogenesis in salmonids, whereas GTH II is predominant at the time of spermiation and ovulation. GTH I and GTH II are found in distinctly separate cells. In trout, GTH I is expressed first in ontogeny, whereas GTH II cells appear coincident with the onset of spermatogenesis and vitellogenesis, and increase dramatically at the time of final reproductive maturation. Comparison of the amino acid sequences of polypeptides and the base sequences of cDNA revealed that salmon GTH I β is more similar to bovine FSHβ than bovine LHβ and salmon GTH II β shows higher homology to bovine LHβ than to bovine FSHβ. The existence of two pituitary gonadotropins in teleosts as well as tetrapods suggests that the divergence of the GTH gene took place earlier than the time of divergence of teleosts from the main line of evolution leading to tetrapods.  相似文献   

2.
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3.
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.
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.
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4.
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.  相似文献   

5.
Two gonadotropic glycoproteins (PmGTH I and II) were purified by ion-exchange chromatography, gel filtration and preparative SDS-PAGE, from pituitaries of red seabream, a marine teleost which has an asynchronous-type ovary and spawns almost daily during the spawning season. The glycoproteins were composed of distinct subunits and the molecular weights were estimated to be 32 and 38 kDa for PmGTH I and PmGTH II, respectively. Both PmGTH I and II were active in two homologous bioassays: in vitro oocyte maturation and/or in vitro estradiol-17 production assays. These two GTHs were distinct in electrostatic properties, molecular weight, stability and yields from pituitaries during the spawning season. These properties suggest that PmGTH I and II correspond to salmon GTH I and II, respectively.A homologous radioimmunoassay with which to measure PmGTH II was developed using a rabbit antiserum against the subunit of PmGTH II and intact PmGTH II as standards and radioactive competitors. Competition curves for red seabream plasma and pituitary extract were parallel to the standard curve, while PmGTH I had low cross-reactivity (3.1 %) with the antibody. This specific RIA system showed an in vivo LHRHa induced GTH surge in the plasma of female red seabream.  相似文献   

6.
Seawater-adapted female chinook salmon ( Oncorhynchus tshawytscha ) in their second summer were treated with one of three doses of recombinant bovine growth-hormone (rbGH) administered in a continuous fashion by osmotic pump for approximately five weeks. Untreated fish, sham-treated fish and fish treated with the low dose of hormone (0.003 μg/g bwt/day) did not differ significantly in their growth performance over the 10 week course of the experiment. By the end of the treatment period (week 6) the mean increases in fork length and weight for these groups were 7.6% and 38.9%, respectively. A dose-dependent enhancement of growth was observed for the fish treated with the mid (0.016 μg/g bwt/day) and high (0.078 μg/g bwt/day) doses of hormone. After 6 weeks, the fish given the mid dose of rbGH had gained 9.6% in fork length and 45.3% in weight, while the fish treated with the high dose of rbGH had gained 13.9% in fork length and 52.9% in weight. A significant increase in specific growth rate was observed for these latter two groups during the treatment period while condition factor declined in response to the high dose of rbGH. This experiment indicates that chronic administration of growth-hormone to chinook salmon can result in a dose dependent elevation of growth rate. Results are discussed in relation to previous attempts to accelerate the growth of Pacific salmon with growth-hormone and the potential for using recombinant growth-hormone to improve the production characteristics of salmon being cultured in seawater.  相似文献   

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8.
The objective of this investigation was to cryopreserve sexually immature gonads of rainbow trout (RBT). Gonads from juvenile male or female RBT were cryopreserved and stored in liquid nitrogen. Individual testes were thawed and surgically transplanted to isogenic male trout; individual ovaries were autografted. Four weeks following transplantation, all male fish were injected with salmon pituitary extract for 8–10 weeks. The transplanted testis was present in 5 of 8 animals in replicate 1 and in 3 of 6 animals in replicate 2. Fertility of sperm obtained from individual testes in replicate 2 was evaluated by in vitro fertilization. Sperm from the intact control testes had a fertilization rate of 95±3% (mean ± SEM), and sperm from transplanted testes had a fertilization rate of 78±7%. Ovaries that were frozen in liquid nitrogen, thawed and autografted to an ectopic site were not present at five months following surgery.  相似文献   

9.
11-ketotestosterone (OT) is a typical androgen of male teleost fish, but information on the question if it is involved in the feedback regulation of pituitary gonadotropin II (GTH-II) secretion is controversial. We have therefore studied the effects of OT on gonadotropin releasing-hormone (GnRH) stimulated GTH-II secretion in male African catfish Clarias gariepinus). In vivo experiments were carried out with intact and castrated fish. OT plasma levels were increased by implantation of silastic capsules containing 11-ketoandrostenedione (OA) which is converted to OT in both intact and castrated fish. When intact males received OA- or blank-capsules, treatment with salmon gonadotropin releasing-hormone analogue (Des-Gly10-D-Arg6-sGnRH-NEt; 0.2 μg sGnRHa/kg body weight) elevated the plasma GTH-11 levels in both groups. However, the levels were about 2 times higher in blank- than in OA-implanted fish. When castrated fish received either blank-or OA-capsules, sGnRHa treatment led to plasma GTH levels significantly higher than in sham-operated fish. However, there was no difference between the blank- or OA-implanted castrates, though OA implantation led to a restoration of OT plasma levels. This suggests that replacement ofOT is insufficient to reverse castration-induced effects. In vitro experiments were carried out with pituitary tissue fragments using a static culture system. The tissue remained sensitive to sGnRHa (5 × 10?9M) for 4 days after the beginning of incubation. Preincubation of pituitary tissue for 24 hours with 25 ng OT/ml medium (80 nM) completely abolished the stimulatory effect of sGnRHa on GTH-II secretion. Tritiated OT was not metabolized by pituitary tissue during 6 hours of incubation. We conclude that 11-ketotestosterone, a quantitatively prominent and non-aromatizeable circulating androgen participates, at least in part by direct action on the pituitary, in the negative feedback regulation of GnRH-stimulated GTH-II secretion in male African catfish.  相似文献   

10.
Molecular cloning and expression of salmon pituitary hormones   总被引：2，自引：0，他引：2  
A cDNA library was prepared from chinook salmon pituitaries. Growth hormone (GH), prolactin (PRL) and the β subunit of gonadotropin (GTH) genes were screened using synthetic oligonucleotides as probes. Full size cDNA clones coding for these polypeptide hormones were isolated and characterized. The cDNA sequences for PRL and βGTH have been reported earlier from our laboratories. The cDNA clone for GH contains 1148 bp and codes for a preGH of 210 amino acids. The chinook salmon GH, reported in the present investigation, differs from chum salmon GH in only 1 amino acid, and from coho salmon GH in 5 amino acids. Plasmids containing modified nucleotide sequences coding for GH, PRL and βGTH were constructed individually into an expression vector using the heat-inducible λ pL promotor. Mature PRL, GH and unglycosylated βGTH were expressed in the bacteria at elevated temperature.  相似文献   

11.
Gonadotropin-releasing hormone and gonadotropin in goldfish and masu salmon   总被引：1，自引：0，他引：1  
Reproductive activities in vertebrates are regulated by an endocrine system, consisting of the brain-pituitary-gonad axis. In teleosts, gonadotropin-releasing hormone (GnRH) in the brain stimulates gonadotropin (GTH) release in the pituitary gland, but because of lack of the portal vessel, it is not known when and how much GnRH is released for the regulation of GTH release. There are multiple molecular types of GnRH in teleosts and several distinct populations of GnRH neurons in the brain. However, we do not know which types and populations of GnRH neurons regulate reproductive activities. Here we summarize our recent studies on GnRH and GTH in masu salmon Oncorhynchus masou and goldfish Carassius auratus. Immunocytochemistry showed the location and molecular types of GnRH neurons. Salmon (sGnRH) and chicken-II GnRH (cGnRH-II) neuronal fibers were widely distributed in the brain of both masu salmon and goldfish. Only sGnRH fibers were observed in the pituitary of masu salmon, whereas both sGnRH and cGnRH-II fibers were observed in the goldfish pituitary, indicating that species specific GnRH profiles are involved in the regulation of pituitary function in teleosts. A series of experiments in masu salmon and goldfish suggest that among GnRH neuron populations GnRH neurons in the ventral telencephalon and the hypothalamus regulate GTH release, and that GnRH of the terminal nerve origin is not essential to gonadal maturation and ovulation. The biological function of other GnRH neurons remains unkown. Two GTHs appear to be characteristic of teleost; however, regulation of reproduction by these GTHs is a question that remains to be elucidated. In salmonid species, it is proposed that GTH I stimulates early gonadal development, whereas GTH II acts in later stages. When GTH expression was examined in goldfish, both GTH I and II mRNA levels in the pituitary gland showed increases in accordance with gonadal development, unlike the sequential expression of GTH subunits in salmonids. The expression of these GTH subunit mRNAs were affected by water temperature, starvation, and steroid hormones in goldfish, but in what manner these two GTHs regulate gonadal development remains to be clarified.  相似文献   

12.
Levels of two moleculer types of gonadotropin-releasing hormone (GnRH), salmon GnRH (sGnRH) and chicken GnRH–II (cGnRH–II) in the various brain regions and pituitary gland of sockeye salmon (Oncorhynchus nerka) and chum salmon (O. keta) during smoltification and spawning migration, respectively, were measured using specific time-resolved fluoroimmunoassay (TR-FIA) systems. Changes in sGnRH levels in different brain regions tended to be specifically synchronized with serum thyroid hormone or pituitary gonadotropin (GTH) levels during smoltification and spawning migration, respectively. In contrast, cGnRH–II levels did not show such synchronized changes. SGnRH and cGnRH–II in various brain regions might have different roles during smoltification and spawning migration of salmonid fishes.  相似文献   

13.
Methods are described for controlling sex differentiation in the coho salmon, Oncorhynchus kisutch. Eyed eggs and alevins were immersed in solutions of estradiol-17β and 17α-methyltestosterone and fed these steroids in the diet for a period of 10 weeks postswim-up. In seven out of 10 groups that received estradiol all fish resembled normal females when sampled at 4 months posthatch. In fish treated with methyltestosterone at all except the lowest dose (25 and 50 μg/l) the gonads resembled neither normal males nor normal females; these gonads were composed largely of connective tissue with only occasional germ cells. Various proportions of male, female and intersex gonads were observed in groups which received androgen or estrogen only in the diet. The implications of the work for salmon culture include the production of sterile fish and increasing the proportion of female salmon in hatchery populations.  相似文献   

14.
Pituitary gonadotropin (GTH) secreting cells and brain gonadotropin-releasing hormone (GnRH) secreting neurons are known to be subjected to feedback control by gonadal steroid in teleosts. In masu salmon, Oncorhynchus masou, salmon GnRH (sGnRH) neurons in the ventral telencephalon (VT) and the preoptic area (POA) are involved in the control of GTH cells because sGnRH synthesis in these areas is activated with gonadal maturation. In this study, we attempted to clarify mechanisms of feedback control of sGnRH neurons by gonadal steroids. We examined the effects of 17-methyltestosterone (MT) on sGnRH synthesis in yearling and 2-year-old female fish (which were immature during experimentation in May), and the effects of castration on sGnRH synthesis in underyearling precocious male fish in August. sGnRH synthesis in the POA, but not in the VT, was increased by MT administration in 2-year-old females only, indicating higher sensitivity to MT in the preoptic sGnRH neurons. Castration increased sGnRH synthesis in the VT but not in the POA. These results suggest that sGnRH neurons in the VT and those in the POA are differentially regulated by gonadal steroids.  相似文献   

15.
Survival and growth in a saltwater net-pen of sexually immature triploid Atlantic salmon Salmo salar L. brown trout Salmo trutta L. hybrids was comparable to that of immature diploid Atlantic salmon. Following 17 months of freshwater rearing, the experimental fish were individually tagged and transferred to a saltwater net-pen where they were raised communally for 376 days. Initial and final average weights were 158 and 760 g per fish for the diploid Atlantic salmon and 209 and 1010 g per fish for the triploid hybrids; weights for the hybrids were significantly larger in both cases (α= 0.05). Survival from transfer to harvest was 43% for the Atlantic salmon and 48% for the hybrids. Average specific growth rate of fish which survived to harvest was 0.42% day-1 for Atlantic salmon and 0.41% day-1 for hybrids; these values were not significantly different. No significant differences were observed in average condition factor and dress-out percentage between crosses. Average gonadal weights and gonadosomatic indices were not significantly different for male diploid Atlantic salmon, female triploid hybrids and male triploid hybrids, but were significantly greater for female Atlantic salmon.  相似文献   

16.
The effects of acclimation temperature and acute temperature change on the uptake and metabolism of the procarcinogen benzo[a]pyrene (BaP) by gill cells of the gulf toadfish, Opsanus beta, were examined. BaP was rapidly accumulated by isolated gill cells and uptake rates were directly proportional to BaP concentration in the medium (1 to 100 μg/ml). Uptake rates were higher in cells isolated from fish acclimated to 18°C when compared to cells from 28°C acclimated fish at all incubation temperatures. When cells were exposed to BaP at the respective acclimation temperatures of the fish, uptake rates were similar (0.14 ± 0.01 at 18°C and 0.12 ± 0.01 μg BaP/s/10 mg cells at 28°C). This finding is discussed in view of results which showed a partial compensation of membrane fluidity in plasma membranes isolated from fish from the two acclimation temperatures. At higher incubation temperatures, cells from fish acclimated to 18°C metabolized BaP at a greater rate than those at 28°C (49.6 ± 1.92 and 43.0 ± 2.24 μg/g/8h, respectively, at 23°C). Low but detectable activities of common biotransformation enzymes (aryl hydrocarbon hydroxylase, glutathione-S-transferase) and cytochrome P-450 content were found, however, no significant differences were evident between cells from fish acclimated to different temperatures. To whom to address correspondence  相似文献   

17.
Levels of two types of gonadotropin-releasing hormone (salmon GnRH and chicken GnRH-II) in the brain and pituitary, and content of gonadotropin (GTHIβ and IIβ) in the pituitary were measured in male masu salmon from hatching to gonadal maturation for three years in order to clarify the involvement of GnRHs in precocious maturation. Underyearling precocious males were distinguishable in summer of year 1 and were marked by an increased GSI. Spermiation was observed among these individuals thereafter every autumn. Pituitary GTHIβ content in both precocious and immature males, and GTHIIβ content in precocious males showed seasonal fluctuations — high in autumn and low in winter. Pituitary GTHIIβ content was low in immature males. Pituitary sGnRH content in precocious males increased from spring to autumn during the three-year period. sGnRH concentrations in discrete brain areas showed seasonal changes — high during autumn to winter and low in summer. Concentrations in the olfactory bulbs and hypothalamus increased significantly in association with testicular maturation during year 3. sGnRH concentrations in the hypothalamus were significantly higher in precocious males than in immature males; this was possibly due to positive feedback of steroid hormones. cGnRH-II was undetectable in the pituitary and no distinct changes were observed in its concentration in the brain in relation to maturation. The phenomenon of underyearling precocious maturation is considered to be triggered before the onset of early summer. It is suggested that males which mature precociously are larger in size and contain much sGnRH in the pituitary before the outward signs of precocity appear; sGnRH may stimulate GTH II synthesis and induce precocious maturation.  相似文献   

18.
Ceratomyxa shasta is a myxozoan parasite of salmonid fish. In natural communities, distinct genotypes of the parasite are associated with different salmonid hosts. To test the hypothesis that genotypes of C. shasta cause differential mortality, the polychaete host was experimentally infected with different parasite genotypes. Genotype I was obtained from Chinook salmon, Oncorhynchus tshawytscha, and genotype II from either coho salmon, O. kisutch, or rainbow trout, O. mykiss, We then challenged four salmonid strains: Chinook and coho salmon that occur in sympatry with the parasite and allopatric Chinook salmon and rainbow trout. Parasite genotype I caused mortality only in Chinook strains, although mortality in the allopatric strain also occurred from exposure to genotype II. A second experiment demonstrated that genotype II could be separated into two biotypes based on differential mortality in rainbow trout and coho salmon. These differential patterns of mortality as a result of infection by certain genotypes of C. shasta support field observations and suggest a co‐evolutionary relationship between these parasites and their hosts.  相似文献   

19.
Activity of the thyroid gland of the coho salmon,Oncorhynchus kisutch, was assessed by physiological, histological and ultrastructural criteria after treatment with graded doses of bovine thyrotropin (bTSH) in January and March. Average plasma thyroxine (T4) levels increased from about 0.8 ng/ml in saline-injected controls to about 15 ng/ml in fish treated with four intraperitoneal injections of 0.8 lU bTSH. Light-microscope observations of one m-thick sections stained with methylene blue and azure II, showed that bTSH treatment increased epithelial height in both presmolts and smolts. Ultrastructural manifestations of increased activity owing to bTSH treatment were also seen, along with evidence of follicle proliferation. Cytoplasmic organelles and secretory granules increased in numbers with increased dosage of bTSH.  相似文献   

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