Alterations in hepatic microsomal protein levels in rainbow trout fed cyclopropenoid fatty acids analyzed by two-dimensional gel electrophoresis

Two-dimensional gel electrophoresis was used to analyze the effect of dietary cyclopropenoid fatty acids on hepatic microsomal polypeptide distribution patterns. Antibodies against rainbow trout type-LM2 cytochrome P-450 were employed to localize the corresponding polypeptide(s) by immunochemical staining. The LM2 antigen was purified from trout that had been fed beta-naphthoflavone. Microsomes from trout fed beta-naphthoflavone showed a decrease in a cytochrome P-450 polypeptide, detected with antibody against LM2. In contrast, microsomes from control fish contained two distinct cytochrome P-450 polypeptides, differing in their isoelectric points. Cyclopropenoid fatty acid treatment caused a preferential decrease in the additional isozyme seen in control samples. The distribution of concanavalin-A-binding glycopolypeptides was also assessed. Surprisingly, the two P-450 isozymes localized from control microsomes were found to bind concanavalin A. In addition to this, the cyclopropenoid fatty acid treatment generated a shift in a closely related group of microsomal glycopolypeptides, labeled gp80, gp82, gp80(1), and gp82(1). A decrease in the levels of gp80 and gp82 and a corresponding increase in gp80(1) and gp82(1) was observed.     

Alterations in the synthesis of proteins in hepatocytes of rainbow trout fed cyclopropenoid fatty acids

Hepatocytes from rainbow trout reared on a diet containing cyclopropenoid fatty acids were analyzed for alterations in protein composition and synthesis by double label experiments. Both cytosolic and microsomal hepatocyte fractions were investigated. In the cytosolic fraction, the synthesis of proteins in the range of 68,000 to 74,000 daltons were significantly decreased. The identity of these proteins remains uncertain. A pronounced depression in both the mass and apparent synthesis of a 200,000 to 240,000 dalton microsomal protein was also observed. Immunoblotting with antibodies raised against goose acetyl-CoA carboxylase and avidin-peroxidase staining suggest that this protein is acetyl-CoA carboxylase. Moreover, synthesis of this protein as well as mass of the protein in cyclopropenoid fatty acid-fed fish were less than 20% of that found in control fish.     

Plasma oestrogen levels in rainbow trout Salmo gairdneri Richardson

Levels of oestrogens in plasma of mature fall-spawning rainbow trout Salmo gairdneri were determined using a radio-immunoassay. No significant difference was found in oestrogen concentration between the sexes (male =2.5 ng/ml; female =4.4 ng/ml); between individual variability was great. Four blood samplings over a 24-h period via cardiac puncture of males revealed no diel variation or change in estrogen levels due to the stress of bleedings. Although no gonado-somatic index-estrogen relationship could be demonstrated for either sex, there was correlation between oestrogen and androgen levels in the female.     

Lipid mobilization during starvation in the rainbow trout, Salmo gairdneri Richardson, with attention to fatty acids

Rainbow trout, Salmo gairdneri , were fed pelleted food ad libitum for 35 and 50 days. Lipid deposits were determined in the viscera, liver and muscle (epaxial and hypaxial). Most of the lipid accumulated in the viscera, but the lipid content of liver and epaxial muscle also increased. Hypaxial muscle lipid content remained the same throughout the feeding period. Upon starving the fish for 27 and 48 days following 50 days of feeding, visceral lipid contributed most to energy metabolism among the depots investigated. Muscle also contributed a considerable share while the absolute amount derived from the liver was much smaller.
Patterns of fatty acid mobilization during starvation were also investigated. Saturated acids were preferentially mobilized from the viscera, resulting in a rise in the percentage of monoenes and polyunsaturates. In liver, the percentage of saturates remained relatively constant whereas the percentage of monoenes declined and polyunsaturates increased. In muscle, a substantial increase in saturates was caused by a decline in monoenes; polyunsaturate content remained constant. As a result of these shifts in relative fatty acid composition the U/S (unsaturates to saturates) ratio rose in viscera and liver and decreased in muscle. The UI (unsaturation index) responded in essentially the same way. Possible explanations for preferential fatty acid utilization are discussed in terms of energetics and structural relationships.     

Oxygen uptake by rainbow trout blood, Salmo gairdneri

Oxygen uptake rate by rainbow trout red cells was measured in the temperature range 5° to 40° C. Between 5° and 20° C there was a steady increase in rate and at 20° C a maximum rate of 2.5 μl O₂/min/ml red cells occurred, followed by a decline in the rate as the temperature increased. A morphological investigation of the red cell was conducted and possible functions of the nucleus in fish red blood cells are discussed.     

Purification and properties of allelic lactate dehydrogenase isozymes at the B2 locus in rainbow trout,Salmo gairdneri

• 1.1. The homotetrameric lactate dehydrogenase (LDH) isozymes, B₄^2′ and B₄^2″, from homozygous rainbow trout livers have been purified to homogeneity by affinity chromatography using a Sepharose-linked oxamate ligand.
• 2.2. The B₄^2″ isozyme has been found: (a) to have increased K_m (pyr) and K_i (lac) values, (b) to bind pyruvate strongly in the absence of coenzymes with a binding constant of 4.02 × 10⁴ M⁻¹ under conditions where other LDH isozymes showed no binding, and (c) to give a nonlinear noncompetitive lactate product inhibition pattern compared to a normal linear noncompetitive inhibition for the B₄^2′ isozyme.
• 3.3. Different kinetic mechanisms have been proposed for the two isozymes and an adaptive functional significance has been discussed.

     

Sexual maturation in triploid rainbow trout, Salmo gairdneri Richardson

This paper compares some morphological and endocrinological characteristics of diploid and triploid rainbow trout.
Significant differences were found between diploid and triploid females in GSI, condition factor, gut weight, liver weight and percentage dress-out, and between diploid and triploid males in GSI, condition factor and gut weight.
Diploid females had large, well-developed ovaries containing yolk-filled secondary oocytes whereas the triploids had only string-like ovaries containing nests of oogonia. No primary oocytes were present.
All the diploid males produced copious quantities of milt but it was possible to express a thin, watery milt containing motile spermatozoa from only two of the 12 triploid males. Testes weights in triploids were similar to those of diploids but, while the diploid testes were packed with spermatozoa, those of the triploids consisted mainly of spermatocytes and spermatids with few spermatozoa present. Measurements of the heads of spermatozoa revealed that those from triploids were larger and had a wider size range than those from diploids.
Levels of testosterone and 11-ketotestosterone in triploid and diploid males were not significantly different. However, levels of testosterone and 17β-oestradiol in diploid females were considerably higher than those of triploid females.     

Compensatory growth in the rainbow trout, Salmo gairdneri Richardson

The effect of a period of starvation and subsequent refeeding on the weight and length of rainbow trout at different times of the year has been investigated. Fish that have been starved for 3 weeks and then fed for 3 weeks show a weight gain equivalent to or greater than that of fish fed normally for the 6 week period, in four out of the five periods studied. The study provided evidence of the adaptation of the fish to starvation followed by what may be termed compensatory growth once feeding was resumed. The length changes of the fish indicate that the weight gains were due to growth rather than increases in gut fat deposits or increased water uptake.     

Developmental regulation of metallothionein mRNA, zinc and copper levels in rainbow trout, Salmo gairdneri

The metallothionein (MT) gene expression profile was followed in rainbow trout during early embryo development and in liver and gonads during the period of sexual maturation. The hepatic MT mRNA levels increase at the end of sexual maturation in both male and female rainbow trout. Although both isoforms of MT mRNA accumulate in the liver, there is a preferential increase in MT-A in the female liver. Concomitantly with this increase in MT there is a redistribution of zinc and copper to MT. In the juvenile female there is an abundance of MT mRNA in the ovaries. This is correlated to high levels of zinc in the MT fraction upon Sephadex G-75 chromatography. During ovary development the MT mRNA levels and the MT-bound zinc levels drop, with an increase in zinc being bound to high-molecular-mass proteins. At ovulation most of the zinc is found in the membrane portion upon centrifugation. In contrast to the ovaries, there are no apparent changes in either trace metal distribution or MT mRNA levels during testis development. In the developing embryo there is an increase in MT-bound copper at gastrulation. This is accompanied by an increase in both isoforms of MT mRNA. At hatch both the copper and zinc levels increase in the MT fraction, with a concomitant increase in mainly MT-A mRNA. These findings indicate that the variations in MT mRNA levels during development are closely associated with metal regulation.     

The palatability of amino acids and related compounds to rainbow trout, Salmo gairdneri Richardson

Using a behavioural assay, the palatability of urea, taurine, betaine and the 20 common amino acids (L-isomers) to rainbow trout was determined. The trout responded positively only to aqueous solutions of leucine, isoleucine, proline, phenylalanine, tryptophan, methionine, glutamate, arginine, taurine and urea. Proline, the most effective compound, was active at 10 ^-4 m but not 10^-5m. Palatibility was not directly related to test solution pH; concentration and molecular structure apparently played a greater role. There is good, though not perfect, agreement between the behavioural results obtained here and published electrophysiological data on the palatal sensitivity of rainbow trout to these substances.     

Nutrient digestibility and growth responses of rainbow trout (Salmo gairdneri) fed different levels of cassava and rice

Seven groups of fingerling rainbow trout (Salmo gairdneri) were fed for 10 weeks on 0%, 10%, 20% and 30% of cassava or rice in isonitrogenous diets.Optimum growth and food utilisation was at 20% dietary cassava. High fibre content of the control diet did not suppress protein digestibility in this group. Rather, at all levels, protein digestibility was good and remained between 84.4% and 87.5%. However, in the control group, carbohydrate digestibility was very poor. The cassava diet which had the highest digestible energy as carbohydrate produced the best growth performance, food utilization and protein sparing. At the levels studied, the dietary carbohydrates produced no hyperglycaemic effect on the fish. There was no evidence of drastic adverse effects on the tissue and liver composition of the fish receiving these carbohydrates.     

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.