α-Melanocyte-stimulating hormone directly increases the plasma calcitonin level and involves calcium metabolism in goldfish

International Aquatic Research - The effects of α-melanocyte-stimulating hormone (α-MSH) on calcium metabolism were examined with goldfish. The scales on the left side of goldfish bodies...     

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.     

Effects of salinity, dietary level of protein and 17α-methyltestosterone on growth hormone (GH) and prolactin (tPRL177 and tPRL188) levels in the tilapia, Oreochromis mossambicus

In the present study, we examined the long-term effects of environmental salinity, diet (35% and 25% crude protein) and 17-methyltestosterone (MT) on corresponding levels of pituitary and serum growth hormone (GH) and prolactins (tPRL₁₇₇ and tPRL₁₈₈) in the tilapia (Oreochromis mossambicus). We observed no discernible patterns in serum GH that would suggest an effect of salinity, diet or MT. However, serum GH levels in all treatments declined at 1 and 3h after first feeding. Serum tPRL₁₇₇ and tPRL₁₈₈ were significantly higher in freshwater (FW) than in seawater (SW) and levels were significantly affected by dietary protein. tPRL₁₇₇ levels were higher in all groups fed a 35% protein diet, but tPRL₁₈₈ levels were higher only in the groups fed the MT-treated 35% protein diet; only serum tPRL₁₈₈ levels were affected by MT. Moreover, serum tPRL₁₇₇ and tPRL₁₈₈ increased throughout the sampling time-course. Subsequent work using fasted tilapia suggests that first feeding is likely to initiate the post-prandial suppression of serum GH levels. In contrast with the picture observed in blood, pituitary glands of SW animals showed higher levels of GH than FW fish. Pituitary GH was elevated by MT in both FW and SW. We also observed that pituitary tPRL₁₇₇ and tPRL₁₈₈ levels were higher in FW fish than in SW fish; tPRL₁₇₇ and tPRL₁₈₈ levels were elevated by MT only in FW animals. To assess the somatomedin activity of plasma from FW- and SW-reared tilapia, we measured [35S]-sulfate incorporation into ceratobranchial cartilage explants in vitro. Plasma from SW-adapted tilapia showed greater activity in this assay than plasma from FW-reared tilapia, suggesting that the GH-dependent IGF bioactivity of plasma is higher in SW-reared tilapia. Collectively, these studies suggest that the growth-promoting actions of SW rearing and of MT administration in tilapia may be linked to elevations in GH and/or prolactin (tPRL₁₇₇ and tPRL₁₈₈)levels.     

Bioaccumulation of hexachlorocyclohexane,dichlorodiphenyltrichloroethane, and estradiol-17β in catfish and carp during the pre-monsoon season in India

This investigation was performed to monitor hexachlorocyclohexane isomers (HCHs), dichlorodiphenyltrichloroethane (DDT, and its metabolites, refered to as DDTs), plasma levels of estradiol-17β (E2), and the gonadosomatic index (GSI) between sampling sites of unpolluted ponds of Gujartal, Jaunpur (control site) and the polluted rivers Gomti (Jaunpur) and Ganga (Varanasi), which affect the reproductive physiology of some edible catfish and carp during the pre-monsoon season. HCHs and DDTs were measured by gas liquid chromatography (GLC) and hormones by radioimmunoassay (RIA). The results indicated that the level of HCHs and DDTs was very high in both the catfish and the carp captured from the polluted rivers compared with the fish captured from the control site. The GSI and E2 values were lower in both groups of fish when compared to the fish from the control site. The results also indicate that catfish showed greater bioaccumulation of HCHs and DDTs than carp, above the permissible limit, as compared to the fish from the control site. In conclusion, fish from the Gomti and Ganga rivers were highly polluted when compared with fish from the control site, as was evident from high levels of tissue bioaccumulation of HCHs and DDTs and decreased levels of plasma E2, inhibiting the reproductive physiology of these species at the receptor level. The levels exceeded the maximum residue limits (MRL) as recommended by Codex, hence it is suggested that the fish should be avoided for food purposes.     

Modulation of glycoprotein hormone α- and gonadotropin IIβ-subunit mRNA levels in the pituitary gland of mature male African catfish, Clarias gariepinus

The cDNAs encoding the glycoprotein hormone -subunit (GP) and the gonadotropin II-subunit (GTH II) were cloned from the pituitary gland of the African catfish, Clarias gariepinus. Using RNase protection analysis, we studied the steady-state mRNA levels of GP as well as GTH II in the pituitary gland of adult male catfish. Castration of adult male catfish resulted in a significant decrease of GTH II mRNA levels, whereas there was no change in the GP mRNA levels. Treatment of intact males with a single dose of 11-ketotestosterone (11-KT) resulted in dose-dependent increases in mRNA levels of both GP and GTH II. We conclude that 11-KT, a prominent, non-aromatizable teleost androgen, has a stimulatory effect on the pituitary mRNA levels of GP and GTH II of adult male fish.     

Studies on activity, distribution, and zymogram of protease, α-amylase, and lipase in the paddlefish Polyodon spathula

A series of biochemical determination and electrophoretic observations have been conducted to analyze the activities and characteristics of protease, α-amylase, and lipase of paddlefish Polyodon spathula. The results obtained have been compared with those of bighead carp (Aristichthys nobilis) and hybrid sturgeon (Huso dauricus ♀?×?Acipenser schrenki Brandt ♂), in order to increase available knowledge of the physiological characteristics of this sturgeon species and to gain information with regard to its nutrition. Further, a comparative study of enzymatic activity, distribution, and characterization between commercial feed-reared paddlefish (CG) and natural live food-reared (NG) paddlefish was conducted. Results showed that higher proteolytic activity was observed in the pH range 2.5-3.0 and at a pH of 7.0 for paddlefish. Levels of acid protease activity of paddlefish were similar to that of hybrid sturgeon, and significantly higher than that of bighead carp. The inhibition assay of paddlefish showed that the rate of inhibition of tosyl-phenylalanine chloromethyl ketone was approximately 2.6-fold that of tosyl-lysine chloromethyl ketone. There was no significant difference observed for acid protease activity between PG and CG groups, whereas the activity of alkaline protease, α-amylase, and lipase in the PG group were significantly lower than those in the CG group. The substrate sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis further showed that there were certain types of enzymes, especially α-amylase, with similar molecular mass in the paddlefish and hybrid sturgeon. It can be inferred that acid digestion was main mechanism for protein hydrolysis in paddlefish, as reported for other fishes with a stomach. This indicates that the paddlefish requires higher alkaline protease, α-amylase, and lipase activity to digest natural live food.     

Habitat and prey selection of common minke,sei, and Bryde’s whales in mesoscale during summer in the subarctic and transition regions of the western North Pacific

This study represents the first quantitative analysis of the characteristics of the distribution areas and stomach contents of common minke whale Balaenoptera acutorostrata, sei whale B. borealis, and Bryde’s whale B. edeni in relation to oceanographic and prey environments in mid summer in the western North Pacific. Common minke whales were distributed within subarctic regions and the northernmost region of the transitional domain, coinciding with the main habitat of their preferred prey, Pacific saury Cololabis saira. Sei whales were mainly found in the northernmost part of the transition zone and showed prey preference for Japanese anchovy Engraulis japonica, which was significantly more abundant in the main distribution area of the whale than in its adjacent areas. “Hot spots” of Bryde’s whales were found in several regions of the transition zone between the subarctic boundary and the Kuroshio front. This whale species preferred Japanese anchovy as prey, for which the distribution density was significantly higher in the main distribution area of the whale than in the adjacent areas. These results indicate that the summer distributions of Pacific saury and Japanese anchovy greatly influence the distributions of these whale species, suggesting that the whales’ habitat selection is closely related to their prey selection.     

Effects of melatonin,p-chlorophenylalanine,and α-methylparatyrosine on plasma gonadotropin level and ovarian activity in the catfish,Heteropneustes fossilis: A study correlating changes in hypothalamic monoamines

The effects of (ip, 10 injections over 20 days) of melatonin (75 g 100 g^–1 BW), the serotonin (5-HT)-synthesis blocker, para-cholorophenylalanine (p-CPA, 10 mg 100g^{–1 BW}) and the catecholamine-synthesis blocker, -methylparatyrosine (-MPT, 10 mg 100 g^–1 BW) on gonadotropin (GTH) secretion and ovarian activity were studied in ^{Heteropneustes fossilis} during late preparatory to early prespawning (April–May). The treatments resulted in significant reductions of plasma GTH and estradiol-17 levels, the gonadosomatic index, frequency distribution of vitellogenic and postvitellogenic oocytes, and ovarian and serum ³²p-labelled alkali-labile phosphoprotein (a marker of vitellogenic activity). Most of the oocytes were nonvitellogenic or had undergone atretic changes. The hepatic ³²-phosphoprotein content increased significantly over the saline control value. The effects were similar and pronounced in the p-CPA and melatonin-treated groups but were moderate in the -MPT-treated group. Hypothalamic 5-HT content and turnover were significantly inhibited in the p-CPA and melatonin-treated groups but the content and turnover of catecholamines were not. The -MPT treatment decreased significantly the content and turnover of dopamine (DA), noradrenaline (NA), and adrenaline (A) but did not influence the 5-HT content or turnover. These results suggest that 5-HT, NA and A are stimulatory to GTH secretion and that melatonin may act on the serotonergic system to inhibit the pituitary-gonadal axis.to whom correspondence should be addressed.A part of the results was presented at the International Workshop on Pineal gland: Its molecular signals and published as an abstract in Neuroendocrinol. Lett. 14: 399 pp., 1992.     

LH-β and FSH-β mRNA expression in nesting and post-breeding three-spined stickleback, Gasterosteus aculeatus, and effects of castration on expression of LH-β, FSH-β and spiggin mRNA

The mRNA expression of the LH- and FSH- subunits were measured in nesting and post-breeding male three-spined sticklebacks, Gasterosteus aculetaus, as well as in castrated and sham-operated nesting males. Furthermore, expression of an androgen induced kidney protein, spiggin, and 11-ketotestosterone (11KT) levels, were measured in the castrated and sham-operated males. Nesting males had significantly higher levels of both LH- and FSH- mRNA expression compared to post-breeding males. Furthermore, sham-operated males had significantly higher levels of LH- mRNA and spiggin mRNA expression than the castrated fish. Expression of FSH-, on the other hand, did not differ between castrated and sham-operated males. There were strong positive individual correlations between circulating levels of 11KT on the one hand and expressions of LH- and spiggin mRNA, whereas the correlation between 11KT levels and FSH- mRNA was weak. The negative effect of castration on -LH mRNA indicates that gonadal hormones stimulate this expression, whereas this was not the case for -FSH. The observed decline in -LH expression after the end of the breeding season may be the result of cessation of the gonadal stimulation of the pituitary. On the other hand, it is not likely that this can explain the decline in FSH- expression.     

Circulating levels and in vitro production of two maturation-inducing hormones in teleost: 17α,20β-dihydroxy-4-pregnen-3-one and 17α,20β,21-trihydroxy-4-pregnen-3-one, in a daily spawning wrasse, Pseudolabrus japonicus

The female bambooleaf wrasse, Pseudolabrus japonicus, spawns daily during the spawning season, and exhibits a diurnal rhythm of ovarian development. In the present study, we have investigated: (1) circulating levels of 17a, 20-dihydroxy-4-pregnen- 17,20-P) and 17,20,21-trihydroxy-4-pregnen-3-one (20-S) in females sampled at different times of the day during spawning season in captivity, and (2) in vitro production of 17,20-P and 20-S by follicle-enclosed oocytes at seven different develo tal stages. In addition, we developed a microtiter plate enzyme-linked immunosorbent assay (ELISA) for 17,20-P. Serum levels of 17,20-P and 20-S showed similar diurnal changes; substantial increases in these levels occurred around the time of germinal vesicle breakdown (GVBD). In vitro experiments showed that massive production of 17,20-P and 20-S occurred in follicles collected just before or during GVBD. Further, acute decreases in 17,20-P and 20-S production were found in the ovarian follicles just prior to ovulation, suggesting inactivation of the maturation-inducing hormone (MIH). These results, taken together with our previous data on the occurrence of GVBD in vitro, suggest a role for both 17,20-P and 20b-S as MIHs in the bambooleaf wrasse.     

Demonstration of primary cilia and acetylated α-tubulin in fish endothelial,epithelial and fibroblast cell lines

Primary cilia (PC) were demonstrated for the first time in fish endothelial, epithelial and fibroblast cell lines through immunofluorescence staining with the monoclonal antibody, 6-11B-1, against acetylated α-tubulin. The study was carried out with eight recently developed cell lines from the walleye, Sander vitreus (Mitchill). These were three fibroblast-like cell lines, WE-cfin11f, WE-skin11f and WE-liver3 from, respectively, the caudal fin, skin and liver, and three epithelial-like cell lines, WE-cfin11e, WE-spleen6 and WErpe from, respectively, the caudal fin, spleen and retina. Also, endothelial-like WEBA from the bulbus arteriosus and glial-like WE-brain5 from the brain were used. Immunocytochemistry revealed strong staining for acetylated α-tubulin in mitotic spindles and midbodies for all cell lines, and in PC for all cell lines except WE-skin11f. Staining of cytoplasmic microtubules (fibrils) was absent in three cell lines, including WEBA, but present in the others, especially WE-skin11f, which might have obscured PC detection in these cells. Tubacin, an inhibitor of histone deacetylase 6, induced cytoplasmic fibrils in WEBA and the intensity of their staining in WE-cfin11f. These results suggest that the cell lines might differ in their deacetylase activities, making them useful for studying this tubulin modification in teleosts, as well as for studying PC.     

Growth Responses,Tissue Composition,and Liver Enzyme Changes in Juvenile Sunshine Bass,Morone chrysops × M. saxatilis,Associated with Dietary Protein and Lipid Level

ABSTRACT

Four isocaloric, semi-purified diets containing low (30%) or high (50%) protein with either 12 or 18% lipid were fed at 4% of body weight to juvenile sunshine bass, Morone chrysops ×M. saxatilis, for 12 weeks (initial weight = 3.4 g). Fish fed the high protein diets gained significantly more weight and had better feed conversions than fish fed low protein diets. There were no significant differences in muscle or total body moisture and lipid or in intraperitoneal fat associated with either dietary protein or lipid levels. However, muscle protein and liver lipid increased significantly at high protein and lipid levels, respectively. Liver alanine amino transferase activities increased significantly at the high dietary protein and lipid levels, indicating that excess dietary protein was being used for non-protein processes. Fruc-tose-1,6-diphosphatase and pyruvate kinase activities indicated that excess dietary protein was not being used to synthesize glucose or glyco-gen. Malic enzyme activities decreased in response to increased dietary lipid, but did not respond to changes in dietary protein, indicating that excess protein was not being used for lipid synthesis. It is hypothesized the excess dietary protein is used for energy production.     

Effects of acute hypoxia and reoxygenation on oxygen sensors,respiratory metabolism,oxidative stress,and apoptosis in hybrid yellow catfish “Huangyou-1”

Fish Physiology and Biochemistry - The regulation mechanism of the hybrid yellow catfish “Huangyou-1” was assessed under conditions of hypoxia and reoxygenation by examination of oxygen...     

Influence of 11-ketotestosterone, 17β-estradiol, and 3,5,3′-triiodo-L-thyronine on distribution and metabolism of carotenoids in Arctic charr, Salvelinus alpinus L.*

This investigation examines the influence of implants containing 11-ketotestosterone (11KT), 17-estradiol (E₂), and 3,5,3-triiodo-l-thyronine (T₃) on astaxanthin metabolism in sexually immature individually tagged Arctic charr. The fish (initial average weight 427 g) were maintained in freshwater for 40 days, and weekly implanted intraperitoneally with oil-based injections containing either 11 KT, E₂ or T₃ at levels of 0.1, 1.0 and 0.1 mg (100 g body weight (BW))^–1, respectively. The control fish were given the oil medium alone (0.2 ml 100 g BW^–1). The diet contained ca. 50 mg astaxanthin kg^–1. Carotenoid composition was monitored in plasma, fillet, liver and skin, and 11 KT, E₂ and testosterone (T) levels in plasma. All hormone treatments reduced plasma T compared to the control. E₂-treated fish had a higher (p<0.05) hepatosomatic index (HSI) than the other treatments. Hormone treatment did not influence gonadosomatic index (GSI). T₃ administration induced a silvery skin appearance. The fillet and plasma carotenoid content decreased during the experiment. 11 KT implantation reduced astaxanthin and idoxanthin concentrations of plasma and fillets, and increased the amount in liver and skin, compared to the other treatments. The relative proportion of astaxanthin to idoxanthin was higher in the control fish and T₃ implanted fish, than in fish implanted with 11 KT or E₂ (p<0.05). Fish treated with E₂ had the highest skin carotenoid concentration. Male fish had significantly higher carotenoid content in plasma, fillet and skin than female fish. This study reveals that sex hormones affect carotenoid metabolism and partitioning among body compartments of Arctic charr, effects differently displayed by the sexes.     

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.     

Influence of oral administration of estradiol-17β and testosterone on growth,digestion, food conversion and metabolism in the underyearling red sea bream,Chrysophrys major

Estradiol-17 (E2) administered in the diet to the red sea bream Chrysophrys major did not affect appetite, food conversion efficiency, protein efficiency ratio and specific growth rate. Serum concentrations of total protein, albumin, globulin, vitellogenin, -amino acids, total lipid, free fatty acids, cholesterol and calcium were elevated. The hepatosomatic index was also increased. Activities of hepatic enzymes including lactate dehydrogenase, isocitrate dehydrogenase and glutamate-pyruvate transaminase were higher than found in untreated control fish. Intestinal activity of leucine aminopeptidase was augmented. However, there were no changes in muscle water, protein, lipid and glycogen content. In contrast, testosterone (T) given by the same route increased appetite, food conversion efficiency, protein efficiency ratio and specific growth rate. There were no alterations in serum protein and calcium concentrations but serum glucose, ammonia and triglyceride levels were elevated. Hepatic glycogen content was increased. The activities of hepatic fructose- 1,6-diphosphatase, glucose-6-phosphatase and glycogen synthetase and intestinal activities of alkaline phosphatase and -glutamyltransferase were higher than noted on control fish. The results reveal that estradiol-17 and testosterone exerted different metabolic effects in the red sea bream and they suggest that testosterone exerts its anabolic actions by increasing appetite, food conversion efficiency and activities of digestive enzymes.     

Seasoned Sprat Products’ Acceptance in Estonia and in Thailand

The aim of the study was to describe and evaluate competitiveness of Estonian spice-cured sprat products in Estonia and Thailand. Sample A was saltiest, Sample B had lowest overall spiciness, and Sample C had highest pepper flavor and sour taste intensity. The main drivers of consumer acceptance for the spice-cured sprat products were different: flavor and appearance in Estonia and appearance and odor in Thailand. In Estonia, one cluster of consumers liked the traditional (Sample A) and lightly spiced (Sample B) sprat products, while the other cluster liked the traditional (Sample A) and the marinated (Sample C) sprat products. In Thailand, all samples scored low, but manual clustering indicated that marinated (Sample C) sprat products are most acceptable. The current study showed that spice-cured sprat products in general can be accepted by Thai consumers, especially as part of meals, if further flavor development is carried out with the products.