Histochemical studies on metabolic zonation of the liver in the trout (Salmo gairdneri)

Summary The livers of 26 adult male and female trout were studied histochemically. G6Pase activity was always found to be heterotopically distributed with a constant maximum in the periportal area. In many cases the glycogen content and the activity of phosphorylase predominated in the periportal zone as well. Maximum activity of glucose-6-phosphate-dehydrogenase and malic enzyme, however, could be demonstrated preferentially in the perivenous area. Lactate dehydrogenase, succinate dehydrogenase, alcohol dehydrogenase, acid phosphatase and -glucuronidase were found equally in all liver cells. 3-Hydroxybutyrate dehydrogenase was absent. Thus, the principles of metabolic zonation have been established in trout liver, the architecture of which differs essentially from that of mammals. The course of the terminal afferent and efferent vessels is the decisive factor for the heterotopic localization of functional units rather than the tubular or plate-forming arrangement of the hepatocytes.Supported by Grants from the Deutsche Forschungsgemeinschaft (Sa 127/8-4)Parts of this study were presented as an Inauguraldissertation to the Medical Faculty of the University of Basel     

Hormonal effects on the liver glucose metabolism in rainbow trout (Salmo gairdneri)

1. Glucagon, adrenaline and dibutyril cyclic AMP increased the release of glucose to the medium during incubation of liver slices from rainbow trout (Salmo gairdneri) while insulin had no effect. 2. Glycogen content decreased only slightly after cyclic AMP addition and even increased in the presence of glucagon and adrenaline. Consequently, the release of glucose was due mainly to gluconeogenesis. 3. This is corroborated by the reduction of glucose liberation in presence of alpha-cyanocinnamate, an inhibitor of gluconeogenesis.     

Spermiogenesis in the rainbow trout (Salmo gairdneri)

In an ultrastructural study on the spermiogenesis of the rainbow trout (Salmo gairdneri R.) four spermatogenetic stages were identified. In young round spermatids, the nuclear chromatin was first heterogeneous (euchromatin and heterochromatin). Subsequently, it became more homogeneous and started to condense in the form of coarse granules and fibers and then into fibrils associated in ribbon-like elements which eventually partly fused together. During early spermiogenesis, a juxtanuclear vacuole appeared in the area where the nuclear envelope was specialized due to condensation of material between the two envelopes and a slight accumulation of nuclear material. This area was finally located in the anterior part of spermatids and spermatozoa; it probably plays a role during fertilization. A flagellar rootlet appeared early in spermiogenesis; it may play a role in the attachment of the flagellum to the nucleus since it persisted until the centriolar complex was definitively fixed in the implantation fossa. The flagellum did not display a plasma membrane and was first located in the cytoplasm, but when it was later extruded from the cell, it acquired a membrane. The cytoplasm was rich in ribosomes (free or in small groups) but poor in membranous organelles. The few mitochondria polarized around the centriolar complex were finally organized into an annular mid-piece. The spermatids remained connected by intercellular bridges until the end of spermiogenesis. The complexity of trout spermiogenesis is intermediate between that in poecilids and that in carp and pike, which have very simple spermatozoa. The role of the material from the nucleus and the cytoplasm reaching the Sertoli cell in the control of spermatogenesis has been discussed.     

Apomorphine-induced vomiting in rainbow trout (Salmo gairdneri)

1. The LD50 for a 7-day period following intraperitoneal injection of apomorphine-HCl was calculated to be 158 mg/kg in rainbow trout. 2. Intraperitoneal injection of apomorphine at doses of 60 mg/kg or greater caused vomiting of plastic balls which had been placed in the stomachs of rainbow trout. 3. Apomorphine-induced effects included vomiting, vomiting behavior, toxicity, increased respiration, impaired motor control and equilibrium, and increased aggression. 4. The vomiting control mechanism of trout may be similar to that described in mammals.     

Evidence for glycosyltransferases in trout liver microsomes (Salmo gairdneri).

1. Trout (Salmo gairdneri) serum is rich in glycoproteins which are synthetized in liver. 2. An attempt to localize glycosyltransferases in hepatocytes is described, using cellular fractionation and marker enzyme determination. 3. Galactosyltransferase, mannosyltransferase, N-acetyl-glucosaminyl transferase, glucosyltransferase, sialyltransferase (on exogenous acceptor) are found in a microsomal fraction obtained by centrifugation at 117 X 10(5) g min of the post-mitochondrial supernatant. 4. Mannose is transferred to endogenous lipids and proteins.     

Energy metabolism of swimming trout (Salmo gairdneri)

Summary The oxidation of 1-¹⁴C palmitate, 2-¹⁴C glucose, 1-¹⁴C lactate, 1-¹⁴C alanine, 1-¹⁴C leucine and 1-¹⁴C glutamate, injected via a cannula into the dorsal aorta, was determined in trout, either at rest, or during swimming at 80% of the maximum sustained speed. The oxygen consumption and the excretion rates of¹⁴CO₂ as well as CO₂ were measured.While the oxygen consumption of swimming trout was about twice as high as of resting trout, the oxidation rates of the injected tracers increased by up to 9 time. Despite the increased importance of blood borne substrates, the estimated contribution to total CO₂ production is about 6% for the resting and 17% for the active trout. The majority of the oxidisable substrates must therefore be endogenous.The mobilization and oxidation rates of lactate, palmitate and leucine were particularly increased during swimming. During rest, palmitate and leucine oxidation rates are low. While oxidation rates of alanine and glutamate are intermediate, those of glucose were found to be extremely low, both at rest and during swimming. The measured RQ values for resting and swimming trout were 0.91 and 0.96 respectively, indicating that protein is the major fuel, since glucose oxidation seems of minor importance.Abbreviations and symbols SA specific activity - tt transit time - decay (time) constant - flux (in % of injected dose per hour) - U_crit critical velocity of sustained swimming - TCO₂ total CO₂     

Chromosome polymorphism in the rainbow trout (Salmo gairdneri Richardson)

Chromosome preparations from lymphocyte cultures of 50 rainbow trout were studied. Diploid chromosome numbers of 59, 60, 61, 62 and 63 were found in different individuals in which the arm number (NF) was 104. Intraindividual polymorphism was found at a low level in 25 of the fish. The results suggest that numerous chromosome polymorphisms exist in rainbow trout.     

Aldehyde dehydrogenase activity in brain and liver of the rainbow trout (Salmo gairdneri Richardson)

Aldehyde dehydrogenase (ALDH) activity was measured in brain and liver of rainbow trout by using 3,4-dihydroxyphenylacetaldehyde (DOPAL, the biogenic aldehyde derived from dopamine) as the substrate. The amount of the corresponding acid produced was quantified by high-performance liquid chromatography with electrochemical detection. Both in brain and liver, the ALDH activity showed a high affinity for the substrate with an apparent Km of 3.7 microM in brain and 2.4 microM in liver. The kinetic experiments with brain ALDH also indicated the presence of an isozyme with a low affinity for DOPAL with a Km around 150 microM. The Vmax of the liver ALDH activity varied between 179 and 536 nmol/min.g, i.e., about 25-75 times higher than that of the low-Km activity in brain. The ALDH activity showed a maximum around pH 8.5, it was stimulated by Mg2+, and disulfiram was found to be a potent inhibitor of the enzyme. The results suggested that the majority of the ALDH activity was located in mitochondria (60-70% with regard to the brain and 70-80% with regard to the liver), while the remaining activity appeared to be cytosolic in both organs. No microsomal ALDH activity could be found.     

Crystallization of the rainbow trout (Salmo gairdneri) haemoglobin IV

Crystals of rainbow trout (Salmo gairdneri) haemoglobin IV were grown in mini batches from a solution of ammonium sulphate. Large single crystals grew over five days and were up to 2 mm in length. X-ray diffraction experiments indicated a space group of C222(1) with unit cell dimensions of a = 85.3 A, b = 94.6 A and c = 105.7 A. The crystals diffract to better than 2.5 A but exhibit some mosaicity along the c axis.     

Endocytosis of galactose-terminated glycoproteins by isolated liver cells of the rainbow trout (Salmo gairdneri)

Intravenously injected 125I-labeled galactose-terminated glycoproteins were mainly recovered in the liver of the rainbow trout. After injection of [14C]sucrose-labeled asialofetuin, the liver cells were isolated and separated by differential centrifugation. The radioactivity was located in the parenchymal cells. Uptake of asialoglycoproteins in liver cells was inhibited by EGTA, lactose and excess unlabeled ligand. Degradation was inhibited by ammonium chloride, suggesting a lysosomal process. Internalization of 125I-asialoglycoproteins was demonstrated by removing receptor-bound ligand with EGTA at different time points during the incubation. The cellular uptake occurred even at 0 degree C.     

Rainbow trout (Salmo gairdneri) stocking and Contracaecum spp

A stocking program with rainbow trout (Salmo gairdneri) at High Rock Lake, Manitoba failed due to infections with large numbers of Contracaecum spp. larvae. Nematode larvae in the intestinal tract, body cavity and musculature made the fish unmarketable. A combination of experimental infections of rainbow trout and pelicans (Pelecanus erythrorhynchos), observations on the behavior of fish-eating birds, and numbers of larval Contracaecum spp. in minnow species led to the following conclusions. The introduction of rainbow trout attracted large numbers of fish-eating birds, particularly pelicans. Concurrent predation by rainbow trout on fathead minnows (Pimephales promelas), five-spined sticklebacks (Culaea inconstans), and nine-spined sticklebacks (Pungitius pungitius), concentrated the parasites. The combined increase in densities of the introduced fish host and fish-eating birds, and the short life cycle of the parasite, increased the numbers of parasites in rainbow trout over a season and in the indigenous minnow species between years. Numbers of larvae in the indigenous minnow species declined when stocking of rainbow trout was stopped and use of the lake by fish-eating birds, particularly pelicans, returned to normal levels.     

Cadmium-binding protein (CdBP) in the isolated perfused liver of rainbow trout, Salmo gairdneri Richardson

An isolated perfused liver preparation was used to demonstrate the production of Cadmium Binding Protein (CdBP) by rainbow trout liver in vitro during perfusion with saline media containing cadmium. Synthesis appears to be both time and concentration dependent and the CdBP has the same characteristics as CdBP produced in vivo.     

Ultrastructure of the physostomatous swimbladder of rainbow trout (Salmo gairdneri)

Summary Rainbow trout swimbladder epithelium consists of non-ciliated and ciliated cells in the ratio of greater than 21. Non-ciliated cells contain vesicles filled with a mucus-like material and similar material is found lining the surface of the swimbladder lumen. Morphological evidence for discharge of the vesicle contents was obtained. In addition, nonciliated cells contain osmiophilic lamellar bodies which resemble the cytosomes of lung alveolar cells of air-breathing vertebrates. The non-ciliated cells do not appear to be involved in a process of active gas secretion.Supported by a research grant from the American Cancer Society, Oregon Division, Inc.     

Some blood parameters of the rainbow trout (Salmo gairdneri Richardson)

Fifty rainbow trout of the Kamloops strain were examined for 12 haematological parameters: erythrocyte sedimentation rate, haemoglobin, packed cell volume, erythrocyte count, erythrocyte diameters, mean cell volume, mean cell haemoglobin, mean cell haemoglobin concentration, leukocyte count, differential leukocyte count, plasma total protein and plasma glucose concentration. The fish had been held under known environmental and dietetic conditions, and at the time of sampling were 14 months old. The majority of results for erythrocyte sedimentation rate, haemoglobin, packed cell volume, erythrocyte count, erythrocyte diameters, total protein and differential leukocyte count fell within narrow ranges. The total leukocyte counts and glucose levels were more widely spread. The results are discussed and compared with those already published for Idaho and Shasta strains. It is impossible to say whether the differences that were observed between Kamloops and these other varieties were due to strain alone, since other variables were present. Some problems associated with establishing normal ranges for these parameters in fish are discussed.     

Seasonal changes in reproductive function of the rainbow trout (Salmo gairdneri)

The sequential changes in serum oestradiol 17β (measured by specific radioimmunoassay), vitellogenin (measured by phosphoprotein phosphorus content) and total calcium (measured by fluorimetry) in male and female rainbow trout under a simulated natural photoperiod cycle were investigated. Resting levels of 130 pg/ml serum oestradiol in both male and female fish were found in April and May. In the female, levels reached a peak of 4800 pg/ml in October, and almost returned to resting levels just prior to spawning in mid-January. No significant change from resting levels was seen in the male fish.
In both male and female fish levels of 25 μg/ml serum phosphoprotein phosphorus and 10–14 mg % total serum calcium were found from April to July. Coincident with the rise in oestradiol 17β, in the females serum levels of phospho-protein phosphorus and total calcium increased to 400 uβg/ml and 58 mg% respectively just prior to spawning. In the males no significant change in either of these values was observed throughout the cycle. These results strongly support the hypothesis that photoperiod is the major factor in the environmental control of reproductive activity in the rainbow trout.     

Some blood parameters of the rainbow trout (Salmo gairdneri Richardson)

Some of the blood constituents of male, female and immature rainbow trout of the Shasta variety, maintained in known environmental and dietetic conditions, were examined and the results statistically treated. The parameters for male and female trout were similar, only the erythrocyte count exhibiting a significant difference. Large but expected differences were evident between the parameters for mature and immature fish.