Development of a protein binding assay for teleost insulin-like growth factor (IGF)-like: relationships between growth hormone (GH) and IGF-like in the blood of rainbow trout (Oncorhynchus mykiss)

Using rainbow trout plasma protein (IGF-BP) which specifically binds human insulin-like growth factor (IGF) (Niu and Le Bail 1993), we have developed an assay to measure plasma IGF-like levels in different teleost species. Before the assay and to prevent interference by IGF-BP, IGF-like was extracted from all samples, using SP Sephadex C-25 in acidic conditions. After this treatment, contamination of the IGF fraction by IGF-BP which was estimated by binding assay, was approximately 5%, and was not detectable by western ligand blot. Human IGF-I was used as standard and labelled hormone. Sensitivity of the assay was 0.15–0.40 ng/ml (ED90) and ED50 was 1–3 ng/ml. hIGF-II crossreaction was partial and no significant displacement was observed with Insulin from different species or with other hormones. Inhibition curves were obtained with plasma IGF fractions (but not with tissue extracts) from teleost and mammals and are parallel to the standard curve. These results suggest that the protein binding assay can quantify an IGF-like factor in the plasma of teleost and that the binding sites of IGF are well conserved during vertebrate evolution. Using this IGF binding assay, IGF-like was measured in parallel with growth hormone (GH) in plasma from young rainbow trout killed every 1.5h throughout one day. The daily profiles for both hormones, which appear pulsatile, are similar. A significant correlation was observed between GH levels and IGF-like levels with a 1.5h delay. Analogous observations were obtained in individual catheterized adult rainbow trout. Although plasma GH levels differ greatly between fish, less variability is found with IGF-like. In a third experiment, rainbow trout were starved or submitted to bovine GH treatment for four weeks. Starved fish, in which plasma GH levels increased, had plasma IGF-like level significantly lower than in fed fish. In bGH injected fish, plasma IGF-like level was significantly higher than in non-injected fish. These results suggest that, as in mammals, IGF-like secretion depends on plasma GH level and could be modulated by the nutritional status of fish.     

Extrapituitary gonadotropin-releasing hormone (GnRH) binding sites in goldfish

In teleosts, as in other vertebrates, the secretion of pituitary gonadotropin (GTH) is mediated by the hypothalamic decapeptide, gonadotropin-releasing hormone (GnRH). Recent findings in teleosts indicate that GnRH receptors are not restricted to the pituitary gonadotropes and are also associated with somatotropes as well as being present in a number of other tissues. In the present study, we provide novel information on GnRH binding in a number of extrapituitary tissues in goldfish. However, we do not intend to provide full characterization of GnRH binding sites in various extrapituitary tissues in goldfish as this would clearly be outside the scope of this paper. In this study we examined GnRH binding in a number of extrapituitary tissues in goldfish and observed specific binding in ovary, testis, brain, liver and kidney. No specific GnRH binding was observed in muscle, skin, gut, gill and heart. In general, the present findings together with the results of other studies carried out in our laboratory demonstrate that mature goldfish ovary and testis contain two classes of GnRH binding sites, high affinity/low capacity and low affinity/high capacity sites with binding characteristics similar to those of the pituitary GnRH receptors. The brain of goldfish was also found to contain two classes of GnRH binding sites, a super-high affinity/low capacity and a low affinity/high capacity sites. Furthermore, study of goldfish liver and kidney demonstrated the presence of a single class of GnRH binding sites with characteristics different from those of pituitary, ovary, testis and brain. Overall, it is evident that goldfish contains a family of GnRH binding sites which can be classified into four groups based on binding affinities: 1) A class of high affinity binding sites present in the pituitary, ovary and testis, 2) a class of super high affinity sites so far only detected in the brain, 3) a class of intermediate-affinity GnRH binding sites in the liver and kidney, and 4) a class of low affinity binding sites present in all the tissues containing specific GnRH binding sites except for liver and kidney.     

Biochemical characterization of growth hormone receptor in rainbow trout (Oncorhynchus mykiss) before and after purification

The aim of this work was to verify if, compared to mammals, the lower molecular weight of GH-R previously reported in salmonid is real or due to the experimental process. For this purpose, we compared the apparent molecular weight of GH-R, obtained by SDS-PAGE after cross-linking with ⁵⁰ I-rtGH, obtained from rainbow trout crude liver membrane preparation, incubated in different buffers with those obtained after purification with affinity chromatography.Using crude liver membrane preparation, two specific bands of ⁵⁰ I-rtGH-protein complex were observed: the major one corresponds to a MW of 70 kDa and the minor one to 45 kDa. However, the pattern of electrophoresis varied according to the different incubation buffers tested. Digestion of the cross-linked complex with -galactosidase and phospholipase did not significantly modify the position of bands, whilst N-glycosidase F induced a large smear including 4 more pronounced bands (50, 65, 97 and > 130 kDa), the heavier band corresponding to the most intensive signal.GH receptors were purified using solubilisation and affinity chromatography. The yield of the liver GH-R from crude liver membrane preparation by the solubilization technique was optimized (48%) using Triton 1% for 1 h (12°C ). Specific binding sites in the solubilized membrane proteins were saturable when incubated with increasing ⁵⁰ I-rtGH concentrations, and revealed a high affinity constant (Ka=0.7×10⁹ M^–1). After affinity chromatography, specific binding activity was increased 64,000 fold. However, the purity of the preparation was partial and the purification yield was very low (about 0.3%). This enriched fraction, analysed by SDS-PAGE after cross-linking, showed a very intense band (about 63 kDa) which disappeared with an excess of cold rtGH.These results suggest that the lower molecular weight observed in salmonid (41 kDa), compared to mamals, is not due to the experimental process. The significance of GH-R size difference between salmonids and mammals is discussed.     

兰州鲇生长激素(GH)基因的克隆及序列分析

利用长片段PCR及T-A克隆技术,从兰州鲇(Silurus lanzhouensis)基因组DNA中首次克隆兰州鲇GH基因全序列并提交至Genbank获得登录号KM215221,同时,使用DNAstar等生物信息学软件进行序列分析研究。兰州鲇GH基因全长2 067 bp,由5个外显子和4个内含子组成,完整编码序列(Complete coding sequence,CDS)为603 bp。5个外显子大小分别为10、140、117、132和204 bp;4个内含子大小分别为229、103、565和103 bp;内含子与外显子的连接区序列遵循基因组成规则。编码区编码1条由200个氨基酸残基组成的蛋白质多肽。5个科8种鲶形目鱼类生长激素基因编码区序列同源性比较表明,它们之间GH基因编码区序列有较高的同源性,平均达到了92.4%,其中,兰州鲇和南方大口鲇之间的同源性最高,为99.5%,兰州鲇和革胡子鲇之间的同源性最低,为90.0%,并构建了系统发育进化树。同时也表明,GH基因编码区序列具有较高的保守性,系统发育研究中具有较高的参考价值。     

Gonadotropin-releasing hormone in two perciform fishes: Micropogonias furnieri and Pagrus pagrus

Gonadotropin-releasing hormone (GnRH) molecular variants were characterized by gradient reverse phase high performance liquid chromatography (RP)-HPLC) from brain extracts of two perciforms with economic importance for Argentina and Uruguay. RP-HPLC fractions were tested in radioimmunoassays (RIAs) with both poly-specific and specific antisera. Both species showed the presence of the same three molecular forms, immunologically and chromatographically indistinguishable from sbGnRH, cGnRH-II and sGnRH. This study supports the hypothesis that their expression is a common pattern in perciforms.     

Dynamics of insulin and insulin-like growth factor-I (IGF-I) ovarian receptors during maturation in the brown trout (Salmo trutta)

In salmonid fishes, the role of insulin and insulin-like growth factor-I (IGF-I) in the regulation of ovarian function is not well understood. Recently, we reported that isolated follicular layers of the preovulatory ovarian follicle of coho salmon (Oncorhynchus kisutch) have specific receptors for insulin and IGF-I and that IGF-I modulates steroid production in the follicular layers. In the present study we have investigated the structural and functional characteristics of insulin and IGF-I receptors in the ovary of brown trout (Salmo trutta) and the changes in insulin and IGF-I binding throughout the reproductive cycle of this species. The specific binding for IGF-I was 8- to 15-fold higher than the specific binding for insulin. IGF-I receptors were also more specific than insulin receptors because unlabeled insulin displaced bound radiolabeled insulin at concentrations 40- to 80-fold lower than unlabeled IGF-I; whereas, unlabeled IGF-I displaced bound radiolabeled IGF-I at concentrations 4000- to 8000-fold lower than unlabeled insulin. Insulin and IGF-I receptors from the brown trout ovary were composed of 120 kDa -subunits and 90 kDa -subunits, which underwent autophosphorylation in a concentration-dependent manner. Receptor tyrosine kinase activity was also stimulated in a concentration-dependent manner by insulin and IGF-I. When ovarian insulin and IGF-I binding was determined from mid-vitellogenesis (March) until ovulation (November), maximal binding for both peptides was detected in mid-vitellogenesis and gradually decreased until the end of vitellogenesis (August). In the preovulatory period (October), a small increase of insulin and IGF-I binding was observed. After ovulation, insulin binding was no longer detectable and IGF-I binding was very low. These results suggest that insulin and IGF-I receptors in the salmonid ovary follow the structural pattern described in other vertebrate species and that insulin and IGF-I could be involved in the regulation of ovarian function during reproductive stages other than the preovulatory period.     

Capture and handling stress affects the endocrine and ovulatory response to exogenous hormone treatment in snapper, Pagrus auratus (Bloch & Schneider)

Sexually mature female hatchery‐reared snapper, Pagrus auratus (Bloch & Schneider) were captured from sea cages by handline and injected at first capture (control) or 24 h after capture, transport and subsequent confinement (delayed injection) with either saline, luteinizing hormone releasing hormone analogue, human chorionic gonadotropin, or 17α‐hydroxyprogesterone. Blood was sampled before hormone treatment and again after 168 h, and fish were checked daily for ovulation. Plasma levels of 17β‐estradiol (E2), testosterone (T), 17α, 20β dihydroxy‐4‐pregnen‐3‐one (17, 20βP) and cortisol were determined by radioimmunoassay. The ovulatory response was assessed from the proportion of fish ovulating, ovulation volume, egg quality and fertility. A delay in injection resulted in significantly lower plasma E2 and T levels in response to hormone treatment, smaller ovulation volumes, and poorer egg quality than in control fish. The results are consistent with the generally inhibitory effects of stress on reproduction in fish, and confirm the requirement to treat fish with hormones designed to induce ovulation, as soon as possible after capture and disturbance.     

Steroid hormone secretion by testicular tissue from African catfish,Clarias gariepinus,in primary culture: identification and quantification by gas chromatography — mass spectrometry

With the final aim of identifying the testicular steroids involved in the feedback mechanism of the hypothalamus-pituitary-gonad axis, steroid secretion by the testis of African catfish, Clarias gariepinus, was studied in vitro, by means of gas chromatography followed by mass spectrometry. Testicular fragments of sexually mature catfish raised in captivity were incubated in L-15 medium with and without catfish pituitary extract (cfPE). Without adding cfPE, 22 steroids could be identified, amongst which 11β-hydroxyandrostenedione, 11β-hydroxytestosterone, 11-ketotestosterone and 11-ketoandrostenedione were dominating. After incubation in the presence of cfPE, the concentrations of the four 11-oxygenated steroids were increased about 4-fold. The amounts of pregnane derivatives in the incubation medium showed the largest increases in the presence of cfPE. 5β-Pregnane-triol levels, for example, were 60-fold higher than in the medium from control incubations. The secretion of 5α- and 5β-androstanes was also stimulated by cfPE. The stimulation was not equal for all steroids, indicating that cfPE not only stimulates total steroidogenesis by increasing the availability of cholesterol, but also by influencing specific steroid converting enzyme activities. Part of this work was presented at the IV^th International Symposium on the Reproductive Physiology of Fish, 7–12 July 1991, Norwich, U.K.     

Endocrine growth regulation of adult Atlantic salmon in seawater: The effects of light regime on plasma growth hormone, insulin-like growth factor-I, and insulin levels

Plasma growth hormone (GH), insulin-like growth factor-I (IGF-I), and insulin were measured in two groups of Salmo salar L. during a one-year study. The fish were reared under either a simulated natural photoperiod (SNP) from January to December or a regime of continuous light from January to June, followed by SNP until December (LL/SNP). Plasma GH levels during spring were low, and lower in the LL/SNP fish (< 0.9 ng ml^− 1) than in the SNP fish (> 1.9 ng ml^− 1), although the LL/SNP grew better (0.8% per day) than the SNP fish (0.5% per day). Plasma IGF-I levels increased transiently from January (64.7 ng ml^− 1) to maximum in late September in the LL/SNP (85.8 ng ml^− 1) and in November in the SNP group (87.3 ng ml^− 1). The ratio GH:IGF-I was lower in the LL/SNP group during spring when this group grew better than the SNP group.     

Steroidogenesis by ovaries and testes of the European catfish,the wels (Silurus glanis), in vitro

Testosterone, 3,17-dihydroxy-5-pregnen-20-one, 17,20-dihydroxy-4-pregnen-3-one (17,20P) and 5-pregnane-3,17,20-triol were identified as the major metabolites of [³H] 17-hydroxyprogesterone in ovarian incubations of the European catfish Silurus glanis. 17,20P and the reduced triol were present only in ovaries from fish primed with carp hypophysial homogenate (chh) while testosterone yields were significantly higher in controls than in treated fish. 11-Ketotestosterone, 11-hydroxytestosterone and 17,20-dihydroxy-4-pregnen-3-one (17,20P) were identified as the major metabolites of [³H]17-hydroxyprogesterone in in vitro incubations of testes of a spermiating catfish. There was no significant production of conjugates or other water soluble metabolites by either sex. The stimulation of plasma 17,20P, 17,20P and 11-hydroxytestosterone by chh in primed but not control males suggests that the role of these steroids in spermiation should be further examined.     

Stable isotope analyses reveal predation on amphibians by a globally invasive fish (Gambusia holbrooki)

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Cortisol and testosterone accumulation in a low pH recirculating aquaculture system for rainbow trout (Oncorhynchus mykiss)

Steroids accumulate in recirculating aquaculture system (RAS), although explanatory factors for such accumulation are still poorly explored. This study investigated the effect of water exchange rate and pH in six replicated RAS on the concentration of the stress hormone cortisol in rainbow trout blood plasma and in the holding water and of the sex steroids testosterone, 11‐ketotestosterone (11‐KT) and 17,20β‐dihydroxypregn‐4‐en‐3‐one (17,20β‐P) over a 70‐day experimental period. Three combinations of water exchange rate and pH were used each treatment, with two replications: (i) high water exchange rate (±1700 L kg^?1 feed) and neutral pH (±7.3), (ii) low water exchange rate (±500 L kg^?1 feed) and neutral pH (±7.3) and (iii) low water exchange rate (±500 L kg^?1 feed) and low pH (±5.8). Plasma cortisol concentrations at day 70 were higher (24.4 ± 9.5 ng mL^?1) for fish kept at low pH when compared to fish kept at neutral pH (12.0 ± 0.1 and 8.7 ± 0.2 ng mL^?1). Water cortisol and testosterone concentrations at day 35 were higher at low pH than at neutral pH, whereas water 11‐KT and 17,20β‐P did not differ among treatments. At day 70, there were no significant differences between low and high pH. These results demonstrate that low pH contributes to increased plasma cortisol concentrations and to its accumulation in water, possibly indicating a stress response to low pH. The higher concentration of testosterone but not of the other sex hormones point to unspecified reproductive effects that need further investigation.     

鸭绿沙塘鳢繁殖习性的观察及性腺发育周期的组织学研究

周年调查了鸭绿沙塘鳢的繁殖习性,观察了其性腺发育的周年组织学变化.试验结果表明,鸭绿沙塘鳢Ⅰ龄性成熟,分批产卵,雄鱼护卵,繁殖旺季为4-6月,补充群体6月开始繁殖.1、2月雌雄鱼的比例为1 : 2,3月至繁殖季节急剧增至1 : 2～3,10-12月约1 : 1.1-5月雌鱼的成熟系数急剧增加,5-6月急剧下降,7月后上升后又逐渐下降.11月至翌年3月,雄鱼的成熟系数最低, 3-4月急剧上升,5-6月略有下降,7-9月又略上升后而降低.破膜后40 d为Ⅰ期精巢; 60～70 d为Ⅱ期,精原细胞呈束排列;约130 d发育至Ⅲ期;约230 d发育至Ⅳ期,精巢内除精原细胞、精母细胞外,还有少量直径3.15～4.20 μm的精子细胞;约360 d(翌年4-6月)为Ⅴ期,有少量长径为2.10 μm的精子.7月Ⅵ期的壶腹内,部分精子排出,但仍有大量残留.破膜后53 d雌鱼的卵巢为Ⅰ期,卵原细胞直径9.1～10.1 μm,同步发育;150 d发育至Ⅱ期,卵母细胞直径83.3～124.3 μm,细胞核直径71.0～97.2 μm,细胞质嗜碱性,核膜内侧有多个核仁,卵母细胞外有一层滤泡细胞,放射膜不明显.约220 d发育至不同步的Ⅲ期,有Ⅰ、Ⅱ期的卵细胞,滤泡膜和放射膜明显.约280 d(翌年3月)卵巢发育至第Ⅳ期,初级卵母细胞体积增大,充满卵黄颗粒,辐射带增厚.滤泡膜、放射膜和细胞膜厚度分别为7.8、13.0、13.2 μm.约370 d(4-5月)发育至Ⅴ期.雌、雄鱼分别以Ⅲ期和Ⅳ期性腺越冬.     

广东鲂的胚后发育

利用组织学方法,对珠江下游野生广东鲂(Megalobrama terminalis)性腺进行了显微观察和研究,描述了其卵巢、精巢的结构特征以及各类型生殖细胞形态。结果显示,广东鲂属于一次性产卵类型,卵巢发育类型为部分同步型。其卵巢为细线或条状,卵巢发育经历卵原细胞期、单层滤泡细胞期、卵黄泡期、卵黄充满期、成熟期和退化期。卵细胞发生经历3个阶段：卵原细胞、初级卵母细胞和次级卵母细胞;卵巢初级卵母细胞中核仁外排现象出现在第2时相末期至第3时相早期;精巢呈线状或直棒状,精巢为叶状型结构。精细胞发生经历5个阶段：精原细胞、初级精母细胞、次级精母细胞、精子细胞及精子;繁殖季节精子充满于精小叶内,精小囊消失。

     

Diet of the Eurasian otter (Lutra lutra) on the River Gradac,Serbia: Predation in a brown trout‐dominated stream

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Effects of insecticide Thiodan® on the morphology and quantification of ovarian follicles in lambaris Astyanax bimaculatus (Linnaeus, 1758) in different treatments

Thiodan^® is an insecticide which can cause morphological changes in fish tissues, dependent on the concentration. To investigate possible morphological changes, this study evaluated the number and diameter of ovarian follicles in lambaris Astyanax bimaculatus exposed to Thiodan^® for 96 h in different treatments at three sub‐lethal concentrations: 1.15, 2.3 and 5.6 μg L^?1 and a control group without Thiodan^®. Females exposed to Thiodan^® were divided into four treatments: (i) without acclimation and without food, (ii) without acclimation and with food, (iii) with acclimation for 10 days and without food; (iv) with acclimation for 10 days and with food. In this study, it was noted that the action of Thiodan^® at sub‐lethal concentrations in a static system did not affect the morphology of follicular development. However, the treatments with acclimation showed lower numbers of primary and secondary follicles when compared to fish from treatments without acclimation. For the groups exposed to Thiodan^®, a greater number of atretic follicles were observed compared to the control groups. The follicular diameter of secondary follicles in fish exposed to Thiodan^® in the treatment with acclimation and with food was highest in the control group (P < 0.05). Furthermore, the mean gonadosomatic (GSI) and hepatosomatic (HSI) indices of treatments with acclimation were lower than those for treatments without acclimation. These data suggest that the fish may be being affected by the pesticide to which the fish were submitted, and may cause impairment to the follicular development.     

The fatty acid compositions of total,neutral and polar lipids in wild and farmed lambari (Astyanax altiparanae) (Garutti & Britski, 2000) broodstock

One way to develop broodstock fish diets is to determine the compositions of wild broodstock tissues and attempt to replicate these compositions in the eggs of farmed fish via dietary manipulation. We collected 30 wild and 30 farmed lambari females that were in the reproductive stage. The extraction and separation of polar and neutral fractions and the saponification and methylation of lipids were performed and analysed in the muscles, livers and ovaries to determine the compositions of the fatty acids via gas chromatography. Regardless of habitat, lambaris mobilize large amounts of fat to the ovaries during the reproductive period, in addition to highly unsaturated fatty acids, such as arachidonic, eicosapentaenoic and docosahexaenoic acids. The wild lambaris were observed to contain higher levels of various fatty acids, including linoleic acid, which is an essential fatty acid. The most abundant fatty acid that was observed in the commercial diet was linoleic acid, which was supplied in all fish farmed tissues. The commercial diet has low AA, EPA and DHA contents, and, higher levels of these fatty acids were recorded in the tissues of farmed lambari, which suggests that this species are able to elongate and desaturate precursors, linoleic and linolenic acids, into highly unsaturated fatty acids.     

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.

     

Effects of growth hormone on plasma ionic regulation,respiration and extracellular acid-base status in trout (Oncorhynchus mykiss) transferred to seawater

The effects of trout recombinant growth hormone (rtGH) treatment (0.25 g g^–1 by intraperitoneal implant) on plasma ionic regulation, extracellular acid-base status and respiration were investigated in freshwater rainbow trout and during a 4-day period after direct transfer into seawater (35 g 1^–1).In freshwater, rtGH treatment resulted in a significant increase in gill (Na⁺, K⁺) ATPase activity and in standard metabolism (MO₂). The latter would mainly result from a higher rate of protein synthesis. Direct transfer from freshwater to seawater induced a decrease in arterial blood pH, far more pronounced in controls than in treated fish. This effect could be regarded in both groups mainly as a metabolic acidosis resulting from extracellular ion composition changes (i.e., an increase higher in chloride than in sodium, more marked in controls than in treated fish). As the rise in PaCO₂, in spite of an increase in ventilatory activity, is more significant in controls than in treated fish, it can be assumed that rtGH treatment lightened the decrease in the gas diffusing capacity of gills induced by transfer to seawater. The initial increase in MO₂ in both controls and treated fish could be the consequence of an increase in energetic cost of ventilation and osmoregulation. Then, in treated fish, the persistent high level of M may indicate a stimulation of intermediary metabolism by rtGH. In addition, the absence in treated fish of an increase in plasma lactate concentration, as observed in controls, would indicate that rtGH attenuated the decrease in O₂ affinity of haemoglobin foreseeable from the metabolic acidosis.This article is dedicated to Professor Claude Peyraud, whose recent death has deeply saddened us. We respectfully pay a tribute to his memory.