Possible origin of extremely high contents of vitamin D3 in some kinds of fish liver

• 1.1. Comparative studies on the possible origin of extremely high contents of vitamin D₃ in some kinds of fish liver were performed.
• 2.2. Neither photochemical formation of vitamin D₃ in fish skin by solar radiation of 7-dehydrocholesterol (7-DHC) nor nonphotochemical enzymatic formation of vitamin D₃ from 7-DHC in fish liver was demonstrated as the origin of vitamin D₃.
• 3.3. On the other hand, when bastard halibuts and carps were farmed from fingerlings to adults with feedstuff's containing vitamin D₂ or D₃, significant amounts of the vitamins were accumulated in the fish liver.
• 4.4. The contents of vitamins D₂ and D₃ in bastard halibut liver increased according to the duration of farming and dose responses of the vitamins in carp liver were observed.
• 5.5. Significant amounts of vitamins D₂ and D₃ in phytoplankton and vitamin D₃ in Zooplankton and small fish were detected.
• 6.6. Therefore, we have concluded that the most probable origin of vitamin D₃ in fish liver is a result of food chains from plankton.

     

Variations in tissue reserves,plasma metabolites and pancreatic hormones during fasting in immature carp (Cyprinus carpio)

• 1.1. Immature carp were subjected to 2-month fasting periods. Mobilization of reserves in liver and muscle, and the energy contribution of each reserve were studied. Changes in plasma glucose, amino acids, insulin and glucagon levels were determined throughout the experiment.
• 2.2. No changes were observed in plasma glucose, insulin or glucagon at 19 days of fasting, but plasma amino acids increased. At 50 days of fasting, both plasma glucagon and amino acids increased, liver glycogen decreased and muscle proteolysis began.
• 3.3. Between 50 and 67 days of fasting, plasma glucose and insulin decreased significantly, while glucagon and amino acids continued to increase. Strong muscular proteolysis was observed while liver glycogen stabilized.
• 4.4. The contribution of each reserve in liver and muscle to energy production throughout fasting is considered.

     

Effect of triton X-100 treatment on the lipid composition,biochemical and functional properties of hake (Merluccius hubbsi) myofibrils purified from pre- and post-spawned fish

• 1.1. The effects of Triton X-100 treatment on the lipid contents and functional properties of hake myofibrils from pre- and post-spawned fish were investigated.
• 2.2. Differences in lipids, biochemical and functional properties of hake myofibrils related to the gonadal condition of fish were observed.
• 3.3. Triton X-100 treatment removed 65% of polar lipids in myofibrils from pre-spawned fish and only 10% in myofibrils from post-spawned fish.
• 4.4. Triton X-100 increased the Hill coefficient to 1.5 in an allosteric type of reaction for the myofibrillar Mg²⁺-ATPase from pre-spawned hake.
• 5.5. The detergent effect observed on the contraction response was greater in myofibrils from prespawned fish than in post-spawned fish.

     

Effects of membrane-perturbing drugs and noradrenalin on cAMP accumulation and red cell water content in carp (Cyprinus carpio) red cells

• 1.1. Carp red cells were treated with drugs that affect the cell membranes. The water content of the cells and the accumulation of cAMP in the cells were measured in normoxia and in hypoxia using non-stimulated and adrenergically stimulated cells.
• 2.2. WGA, DIDS + CCCP and A23187 increased the water content of nonstimulated normoxic cells.
• 3.3. In hypoxia ouabain and DIDS + CCCP increased the water content but cytochalasin B, NPM, DIDS, CCCP and A23187 + CA²⁺ abolished the hypoxia-induced swelling.
• 4.4. Any membrane perturbation induced some cAMP formation, Sophora and Anquilla lectins being most potent.
• 5.5. Also in adrenergically stimulated cells, membrane perturbation generally increased cAMP formation.
• 6.6. However, cAMP accumulation diminished in cells treated with cytochalasin B, CCCP and DIDS + CCCP.
• 7.7. The adrenergic swelling of carp red cells was reduced in normoxia by DIDS. NPM and CCCP increased the adrenergic swelling in normoxia to hypoxic level.
• 8.8. In hypoxia WGA and Anquilla lectin decreased the swelling.

     

Regulation of fish glycolysis—gluconeogenesis: role of fructose 2,6 P2 and PFK-2

• 1.1. Fructose 2,6 P₂ and PFK-2 have a key role in the regulation of glycolysis-gluconeogenesis in fish
• 2.2. PFK-1 and FBPase-1, as in mammals, are the target enzymes for fructose 2,6 P₂, this in turn may be controlled by glucagon and insulin.
• 3.3. PFK-2 from fish liver seems to be a bifunctional enzyme regulated by phosphorylation/dephosphorylation.
• 4.4. Starvation, refeeding, diet composition and anoxia studies provide a general view of the fructose 2,6 P₂ fish system from which the differences between fish and mammal glycolysis-gluconeogenesis may be ascertained.

     

Effect of repetitive cortisol and thyroxine injections on chloride cell number and Na+/K+-ATPase activity in gills of freshwater acclimated rainbow trout,Salmo gairdneri

• 1.1. Freshwater nonanadromous rainbow trout, Salmo gairdneri, were injected three times a week with either saline, 10μg cortisol/g, 1.0μg thyroxine/g or 10μg cortisol/g + 1.0μg thyroxine/g during a period of 28 days (12 injections). A separate group was derived as a subgroup from the thyroxine group on day 14 and received Cortisol + thyroxine from day 14 until day 28 (six injections).
• 2.2. Gill chloride cell number and Na⁺/K⁺-ATPase activity increased by cortisol treatment, the changes being significant on days 7 and 14, respectively.
• 3.3. Thyroxine treatment did not affect gill Na⁺/K⁺-ATPase activity or chloride cell number directly. Neither did it modify the stimulatory effect of cortisol on these parameters.
• 4.4. Muscle water decreased in cortisol-treated fish and increased in thyroxine-treated fish, while no changes were observed in the combined hormone groups.
• 5.5. No changes were observed in plasma chloride in any group during the experiment.
• 6.6. The results demonstrate a putative role of cortisol in stimulating hypo-osmoregulatory mechanisms and suggest that thyroxine is without a direct or a supportive effect for cortisol action.

     

Strontium-calcium concentration ratios in fish otoliths as environmental indicators

• 1.1. External and internal examinations of otoliths in fishes for macrostructure and microstructure has demonstated yearly, daily and population rhythmic patterns.
• 2.2. Chemical analyses (atomic absorption) of otolith carbonate from reared Fundulus heteroclitus for strontium-calcium concentration ratios demonstrated changes in chemistry related to temperature.
• 3.3. Microprobe analyses made it feasible to interpret almost daily changes in temperature to provide the temperature history of an individual fish.
• 4.4. A combination of microprobe analyses and daily increment analyses of otoliths can provide a life history profile for individual fish and can provide information on the environmental history of each fish.
• 5.5. Such information is vital to our understanding of the processes underlying recruitment and growth rates, and would make it possible to link growth and mortality rates to environmental occurrences.

     

The effect of dietary protein level and ration level on excretion of ammonia in common carp,Cyprinus carpio

• 1.1. Nitrogenous excretion in the form of ammonia was determined in common carp of 65.0 ± 8.0 g body weight in metabolism chambers. The fish were fed with 20, 35 and 50% dietary protein at 1, 2 and 3% body weight per day ration level.
• 2.2. Nitrogenous excretion as a percentage of ingested food increased with an increase of dietary protein but decreased with an increase of ration level.
• 3.3. The energy lost in excretion ranged from 4.19% with 20% dietary protein at 3% ration level to 8.74% with 50% dietary at 1% ration level.

     

Biphasic action of growth hormone on intestinal amino acid absorption in striped bass hybrids

• 1.1. Weekly injections of bovine growth hormone (bGH) increased the maximal transport rate of both Na⁺-dependent and Na⁺ -independent l-leucine transport with little effect on the affinity constants in the intestine of striped bass hybrids.
• 2.2. The Na⁺-dependent and the Na⁺-independent transport of the non-metabolizable analog cycloleucine was also stimulated by bGH.
• 3.3. The Na⁺ -dependent active transport was stimulated 2 days after the hormone treatment, while the stimulation of the Na⁺-independent diffusional transport was not observed until after 2 weeks of treatment.
• 4.4. Studies of intestinal morphometry and l-leucine transport using brush border membrane vesicles suggested that bGH affects intestinal amino acid absorption initially by increasing the number of transporters per cell.
• 5.5. This phase is followed by a general increase of the intestinal mass after long-term treatment with the hormone.

     

Hormonal regulation of human apolipoprotein E gene expression in HepG2 cells

• 1.1. Hormonal regulation of apolipoprotein E (apoE) gene expression by insulin and thyroid hormone was studied in a human hepatoma cell line, HepG2.
• 2.2. Changes at the mRNA level, mRNA translation, in vivo synthesis and secretion were monitored.
• 3.3. Both insulin and triiodothyronine were found to have no significant effect on apoE mRNA levels.
• 4.4. Insulin treatment caused an inhibition of: (a) the in vitro translation of endogenous apoE mRNA in a HepG2 cell-free system (25%), and (b) the incorporation of radioactivity into newly-synthesized apoE in an in vivo pulse-chase labeling experiment (32%).
• 5.5. Interestingly, apoE secretion rate was found to be significantly reduced with insulin (84%) suggesting that a major portion of newly-synthesized apoE may be shunted into a degradative pathway.
• 6.6. Using a similar experimental approach, triiodothyronine showed no significant effect on the rate of apoE synthesis or translation (6–15% decrease), however a slight reduction (20%) in secretion rate was shown.
• 7.7. Overall, apoE gene expression does not appear to be influenced by triiodothyronine significantly but is modulated by insulin at the translational and post-translational level.

     

The effects of cortisol administration on intermediary metabolism in teleost fish

• 1.1. This Mini Review deals with the metabolic consequences of administration of the hormone cortisol on proteins, carbohydrates and lipids in teleost fish.
• 2.2. Many effects of administered cortisol on intermediary metabolism in fish have been reported: inhibition of protein synthesis and/or catabolism of tissue protein which result in higher availability of amino acids, induction of gluconeogenesis and of liver aminotransferases, hyperglycemia and glycogen deposition in the liver, induction of gluconeogenic enzymes, liberation of free fatty acids and deposition of liver lipids. All these effects are observed to a greater or less extent. However, the experimental data show that some effects are inconsistent.
• 3.3. Some explanations for the inconsistencies are given.

     

Troponin C of the Antarctic icefish (Champsocephalus gunnari) white muscle

• 1.1. The troponin C (TN-C) from the Antarctic icefish (_{Champsocephalus gunnari}) has been isolated to homogeneity by a procedure involving extraction from acetone powder, DEAE-Sepharose 4B and AcA 54 column chromatography.
• 2.2. The calcium-induced conformational changes of apo TN-C have been studied by absorption difference spectroscopy, circular dichroism and intrinsic fluorescence.
• 3.3. The results indicate that the overall characteristics of icefish TN-C (such as amino acid composition, modifications of helix content and of the microenvironment of aromatic residues as a function of calcium binding) are quite similar to those of rabbit TN-C.
• 4.4. The intrinsic fluorescence properties are close to those reported for pike and carp TN-C.

     

Binding of insulin and insulin-like growth factor I in bovine chromaffin cells in primary culture

• 1.1. Insulin and insulin-like growth factor I (IGF-I) receptors were studied in bovine chromaffin cells isolated from the medulla by collagenase digestion and kept in primary culture.
• 2.2. Specific ¹²⁵I-labelled insulin binding increased with time in culture with no significant change in the dissociation constant, K_d~0.3nM. Insulin was nearly 100-fold more potent than IGF-I in displacing ¹²⁵I-labelled insulin.
• 3.3. Affinity crosslinking and SDS gel electrophoresis revealed increased binding of ¹²⁵I-labelled insulin and ¹²⁵I-IGF-I with time in culture, the densities of the labelling indicating relatively a much higher expression of IGF-I than insulin receptors in the cells. The apparent molecular weight of both the hormone binding subunits were 135,000, suggesting that the insulin and IGF-I receptors in the adrenal medulla are of the peripheral types.
• 4.4. Both receptors thus appeared to be affected by the collagenase treatment but with a subsequent recovery when cells were kept in culture.

     

Erythrocyte supply from the spleen of exercised carp

• 1.1. Weight and hemoglobin content of the spleen of 3-year-old carp under the resting condition showed seasonal change, and those in male fish were larger than those in females at any season.
• 2.2. Weight of the spleen of 3-year-old fish of both sexes (examined in August and October) and immature fish (examined in September) showed a decrease of more than 50% during severe exercise for 60 min. Hemoglobin content per unit body weight of the same fish showed a decrease of more than 80% with the same exercise.
• 3.3. Hematocrit value and hemoglobin concentration of the blood elevated 41 and 25%, respectively, in the 3-year-old male fish, and 20 and 11%, respectively, in the 3-year-old females during the exercise for 60 min.
• 4.4. Fifty percent and 30% of the hematocrit elevations are considered to have been caused by erythrocyte supply from the spleen in the male and female fish, respectively, and 80% of the elevations in the fish of both sexes can be explained by erythrocyte supply, erythrocyte swelling and water shift out of the blood vessel.

     

Renal response to hypoxia in carp,Cyprinus carpio: Changes in glomerular filtration rate,urine and blood properties and plasma catecholamines of carp exposed to hypoxic conditions

• 1.1. Changes in glomerular nitration rate (GFR), urine and blood properties and plasma catecholamines of carp were investigated during and following hypoxia.
• 2.2. GFR and urine flow decreased with increased urinary concentrations of bio-components, except protein, in the course of hypoxia.
• 3.3. Decreases in blood pH, and increases in haematocrit value and plasma K⁺, Ca²⁺, Mg²⁺, inorganic phosphate (P_i), ammonia, lactic acid and catecholamines (CAs) were observed as hypoxia progressed.
• 4.4. Increased GFR and urine flow, and higher values for urinary components, except protein, compared with those of the control were found in the initial post-stress stage.
• 5.5. The possible significance of increased plasma CAs in relation to changes in renal function in hypoxic carp is discussed.

     

Thermal stability of fish myosin

• 1.1. Thermal stability of fish myosin has been studied by using differential scanning calorimetry (DSC) and circular dichroism (CD).
• 2.2. The temperature range of the sharp decrease in α-helical content agreed very closely with that of the endothermic peaks.
• 3.3. There was a high correlation between the enthalpy of denaturation (ΔH) and the decreasing quantity in α-helicity (Δh).
• 4.4. The structure of fish myosins was much more unstable than that of rabbit.
• 5.5. The instability of fish myosins was reflected in its rod moiety.

     

Clastogenic effects of five carcinogenicmutagenic chemicals on the cells of the common carp,cyprinus carpio L.

• 1.1. The cytogenetic effects of various i.p. treatments with five carcinogenic-mutagenic chemicals and three doses for each (aflatoxin Bl, Aroclor 1254, benzidine, benzo[a]pyrene and 20-methylcholanthrene), were investigated in the cells of the common carp, Cyprinus carpio.
• 2.2. Injection with distilled water and corn oil served as the two control groups.
• 3.3. For detecting cytogenetic damage we used two test systems, chromosomal aberrations (CA) in kidney cells and micronucleated erythrocytes (M).
• 4.4. At 48 hr after treatment with the chemicals under investigation, the frequencies of CA and M were clearly increased in a dose-response manner compared to the control groups.

     

Activation and inhibition of insulin receptor autophosphorylation by trypsin treatment of intact H35 cells

• 1.1. Treatment of intact cultured H35 cells with trypsin (1 mg/ml) for 15 min at low temperature (4°C) or for 30 sec at 37°C causes activation of the insulin receptor subsequently isolated from the cells.
• 2.2. Receptor activation was assessed by increased phosphotyrosine content of the β-subunit of the receptor, and increased autophosphorylation using [³²P]-ATP.
• 3.3. Treatment of the cells for 15 min at 37°C however completely abolished insulin binding and all insulin receptor kinase activity.
• 4.4. These data demonstrate that proteolytic damage of the extracellular domain of the insulin receptor can render the receptor kinase inactive and lead to a cell which is unresponsive to insulin.

     

Ultrastructure of maturing fish (Oreochromis niloticus) and snake (Waglerophis merremii) erythroid cells with regard to hemoglobin biosynthesis

• 1.1. Fish and snake immature erythrocytes were submitted to a comparative ultrastructural study, analysing changes in organelles involved in hemoglobin (Hb) biosynthesis.
• 2.2. Iron uptake occurs probably via transferrin, and ferruginous compounds accumulate as siderosomes, taken as iron sources for heme biosynthesis, later on caught by a double lamella.
• 3.3. Mitochondrial membrane of the inner camera differentiates to lamellated bodies that, sucessively, give rise to expansions for ferruginous material and globin chains captation, constituting prehemosomal vesicles, which become condensed vesicles, followed by prohemosomes.
• 4.4. Through an internal membrane rearrangement, prohemosomes change to hemosomes wherein, hypothetically, heme and the globin chains assembly may occur.
• 5.5. In both fish and snake erythroid cells, all stages for hemosomegenesis are similar to the stages found in erythroid cells of other vertebrate species, including humans, except that fish cells often present single organelles of still unknown function, void of internal membrane.
• 6.6. Through electrophoresis of the respective supernatants obtained after osmotical lysis of the organellar fractions, it was shown that fish hemosomes contain three Hb patterns, while snake hemosomes present two patterns.

     

Insulin,glucagon and catecholamine interactions in avian liver explants

• 1.1. A mechanical tissue chopper was used to obtain liver explants (35–75 mg) from 2- to 3-week-old chickens to determine both tissue sensitivity and metabolic effects of isoproterenol, avian insulin and glucagon.
• 2.2. Avian insulin had no effect on lipogenesis; however, lipogenesis was decreased by dibutyryl cyclic AMP. Insulin did not overcome a decrease in lipogenesis caused by catecholamines. Therefore, this control mechanisms is not modulated by insulin.
• 3.3. Preincubation in the presence of glucagon decreased in vitro lipogenesis. Preincubation in the presence of a 19–29 amino acid construct that approximated the radioimmune site for glucagon did not result in a similar effect. Therefore, this site does not relate to the biopotency of the hormone.
• 4.4. A previously noted catecholamine induced decrease in in vitro lipogenesis was verified, showing that points of in vitro regulation are under phosphorylation-dephosphorylation control.
• 5.5. Preincubation of slices (1 hr) with propranolol blocked the inhibition of lipogenesis caused by α and β adrenergic agonists (arterenol or isoproterenol) during a subsequent 2-hr incubation.
• 6.6. Preincubation of slices with either of these agonists decreased lipogenesis even following an extensive washout.
• 7.7. Inhibition could be overcome with propranolol, a β adrenergic antagonist.