Optimal isolation conditions for rainbow trout (Salmo gairdneri) hepatic ribosomes

1. Isolation conditions for rainbow trout (Salmo gairdneri) liver ribosomes were optimized. 2. Optimal initial buffer (Buffer I) concentrations were 250 mM sucrose, 50 mM Tris (pH 7.6, 25 degrees C), 75 mM KCl, and 5 mM MgSO4.7H2O. 3. Optimal concentrations for post-105 supernatant buffer (Buffer III) were 25 mM Tris (pH 7.6, 25 degrees C), 75 mM KCl, and 8 mM MgSO4.7H2O.     

Ultrastructure of the liver of the fingerling rainbow trout Salmo gairdneri Richardson

Portions of the livers of fingerling rainbow trout were studied by light and electron microscopy. The histology, cytology and ultrastructure of mesothelial cells, serosal fibroblasts, hepatocytes, sinusoidal endothelial cells, endothelial cells of central veins and blood cells were described. Mesothelial cells and fibroblasts constituted a very thin capsule. Hepatocytes contained extensive areas of rough surfaced endoplasmic reticulum, consisting mainly of parallel cisternae and pools of glycogen. One or two nuclei and numerous mitochondria occurred in the areas of endoplasmic reticulum, but never in the pools of glycogen. Hepatocyte surface possibilities included hepatocyte to hepatocyte, hepatocyte to bile canaliculus, hepatocyte to space of Disse and hepatocyte to serosa. The trout liver was compared compared to channel catfish liver and to rat liver. Functional implications of the structural features were discussed.     

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.     

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.     

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.     

Thermal acclimatization of hepatic polysubstrate monooxygenase and UDP-glucuronosyltransferase of mature rainbow trout (Salmo gairdneri)

The thermal acclimatization of hepatic cytochrome P-450-dependent polysubstrate monooxygenase (PSMO) and UDP-glucuronosyltransferase of mature rainbow trout (Salmo gairdneri) was studied. The results indicate that the PSMO system, 7-ethoxycoumarin and benzo(a)pyrene as substrates, shows almost ideal acclimatization pattern in autumn during water cooling. The enzyme activities were identical if measurements were carried out at acclimatization (=environmental) temperatures which were 20 degrees C in August and 5 degrees C in November. If a constant incubation temperature (18 degrees C) was used, the PSMO activities were significantly higher in cold (5 degrees C)-acclimatized fish. The acclimatization process could be seen both in specific and total activities. The content of cytochrome P-450 remains at constant level from August to November. In early summer during water warming the PSMO activities increased considerably in both sexes in all incubation conditions. The specific and total UDP-glucuronosyltransferase activities were significantly higher in warm-acclimatized fish both in the autumn and in the spring if the activities were measured at environmental temperature. No differences could be detected if the measurements were carried out at constant experimental temperature (18 degrees C).     

Properties of microsomal acyl-CoA: lysophosphatidylcholine acyltransferase from liver of thermally-acclimated rainbow trout,Salmo gairdneri

Summary Acyl-CoA: lysophosphatidylcholine acyltransferase (LPCAT) (EC 2.3.1.23) activity was assayed in liver microsomes from rainbow trout,Salmo gairdneri, acclimated to 5°C and 20°C to assess its contribution to the temperature-induced restructuring of phospholipid acyl chain composition. The synthesis of phosphatidylcholine (PC) (from lyso-PC) was threefold the synthesis of phosphatidylethanolamine (PE) (from lyso-PE) under similar assay conditions. LPCAT activity (i) displayed an absolute requirement for lysophosphatidylcholine (LPC) and was enhanced by the presence of ATP, MgCl₂ and CoA (which reduced the impact of endogenous acyl-CoA hydrolase activity by regenerating the acyl-CoA substrate) in the assay medium; (ii) remained linear with time up to 30 min; and (iii) increased linearly with microsomal protein concentration up to 0.2 mg/ml for the 20°C assay and 0.4 mg/ml for the 5°C assay. There was no difference in K_m or V_max values due to the acclimation history of the fish, but there were obvious differences due to assay temperature. The apparent K_m values for LPC were 58.54±7.24 M and 12.26±2.14 M when assayed at 5°C and 20°C respectively; values for oleoyl-CoA were 9.11±0.78 M and 1.23±0.25 M under the same assay conditions. Activity was 1.99±0.31 nmol min^–1 mg protein^–1 when assayed at 5°C, and 3.8±0.45 nmol min^–1 mg protein^–1 when assayed at 20°C. These findings indicate that adjustments in the activity of LPCAT play no significant role in the temperature-induced restructuring of PC molecular species composition. However, the marked temperature dependence of the K_m values for LPC and oleoyl CoA suggest that patterns of fatty acid incorporation (i.e. substrate preference) may vary with assay temperature, and in this way LPCAT could contribute to the restructuring response.Abbreviations PC phosphatidylcholine - PE phosphatidylethanolamine - LPCAT acyl-CoA: lysophosphatidylcholine acyltransferase - LPEAT acyl-CoA: lysophosphatidylethanolamine acyltransferase - LPC 1-palmitoyl,2-lysophosphatidylcholine     

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.     

The saccus dorsalis of the rainbow trout,Salmo gairdneri Richardson

The saccus dorsalis of the brain of the rainbow trout, Salmo gairdneri Richardson, has been investigated by means of histological, cytochemical, enzyme-cytochemical, electron microscopical autoradiographical techniques. The saccus dorsalis is a rostro-dorsal evagination of the diencephalic roof, and consists of a partly folded epithelial wall separating the cerebrospinal fluid from the meningeal matrix fluid. The well-developed vascular system around the epithelial wall, consisting of capillaries with different diameters, seems to be part of the pineal vascular system. No structures were found that may be involved in a possible mechanical or nervous blood flow control. The single-layered epithelium consists of highly specialized cells of one specific type. These cells are mainly characterized by infolded basal membranes, long microvilli of a peculiar shape, non-folded lateral membranes bordering intercellular spaces, apical concentrations of elongate and cup-shaped macromitochondria, a basally located rough endoplasmic reticulum, an apically situated smooth endoplasmic reticulum and apical concentrations of micropinocytotic vesicles. Morphological evidence is presented of a multiple function of these cells: (1) fluid secretion, (2) extrusion of low molecular weight organic substances into the ventricular system, (3) uptake of high molecular weight substances, and (4) uptake of low molecular weight organic substances (aminergic neurotransmitters [GABA]) from the cerebrospinal fluid. The significance of light and dark cells is discussed. Indications of a possible innervation of the saccus dorsalis epithelial cells were not observed. The functional significance of the saccus dorsalis (possible analogue of the choroid plexus?) is discussed.     

Glucose transport across the gill of the rainbow trout, Salmo gairdneri

Glucose fluxes across the gills were measured in freshwater-adapted trout (Salmo gairdneri) using an in vitro, perfused-head preparation. A large asymmetry was observed for the primary lamellar pathway, glucose permeability in the serosa-to-mucosa direction being up to 24 times greater than the permeability in the mucosa-to-serosa direction. Chloride cells appeared to possess a maximal rate of transport, or TMG, of 79 mumol/hr per 100 g. Phlorizin, phloretin and, to a lesser extent, harmaline caused an increase in the rate of glucose efflux. The results suggest that the tubulo-vesicular reticulum, into which plasma is introduced under low pressure, may be regarded as a reabsorption site for glucose in a way similar to the nephron proximal tubule. Thus, essential molecules such as glucose are removed while excess or non-essential substances are excreted into the external medium.     

Deviation from morphological intermediacy in interstrain hybrids of rainbow trout,Salmo gairdneri

Synopsis Deviations from morphological intermediacy in six first generation hybrids between three hatchery strains of rainbow trout, raised in a common environment, are reported. Hybrids have higher mean counts of four meristic characters than their maternal parental strain in a significantly greater number of cases (18 out of 24). Furthermore, eight of eleven hybrid indices are not intermediate. These results are discussed in reference to several mechanisms and models proposed to account for observed responses of meristic characters to environmental and genetic influences.     

Synthesis of platelet activating factor by tissues from the rainbow trout, Salmo gairdneri

In mammals, platelet activating factor (PAF, 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) is a lipid mediator with biological activity at concentrations in the subnanomolar range. Although PAF is known to have many activities in mammals, little is known about its synthesis and importance in other vertebrate groups. We demonstrate here the synthesis of PAF from [3H]acetate by slices of trout gill, kidney, liver and spleen. PAF synthesis was stimulated by the calcium ionophore A23187 and was time-dependent. The radiolabeled PAF produced was characterized by TLC, HPLC, derivatization and by saponification and phospholipase A2 hydrolysis. These findings suggest that PAF may be an important mediator in fish.     

Effects of sampling conditions on selected blood variables of rainbow trout, Salmo gairdneri Richardson

The effects of two methods of specimen immobilization (MS 222 anaesthesia and stunning), two types of anticoagulant (EDTA and heparin), two storage temperature ranges (0–2°C and 22–25°C) and four sample storage periods (0, 1, 3, and 24 h) on the haemoglobin, haematocrit, plasma and packed cell sodium, potassium and chloride ion concentrations and packed cell ATP levels of rainbow trout were examined. Stored samples exhibited increases in cell volume, net transfer of sodium and chloride from plasma into cells, net loss of potassium to plasma and rapid depletion of ATP. Room temperature conditions and prolonged storage exacerbated these changes. Use of EDTA, particularly in combination with MS 222, frequently led to haemolysis. Least change in most variables was observed in samples drawn from stunned specimens, treated with heparin and refrigerated before use or preparation for deep cold storage.     

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.     

Cadmium uptake by rainbow trout, Salmo gairdneri eggs and alevins

The uptake of cadmium by eggs and alevins of rainbow trout from water concentrations of between 0.01 and 50.0 mg Cd 1⁻¹ was investigated. The cadmium content of eggs and alevins increased with time and with exposure concentration. Lower cadmium levels were detected in alevins than in eggs. Most of the cadmium (98%) in the eggs was found to be associated with the egg membrane or chorion. This explains the considerable reduction in cadmium concentration observed in alevins after hatching. Alevins hatching from eggs which had been exposed to cadmium survived longer in cadmium than alevins not exposed as eggs. This suggests that the pretreatment of eggs with cadmium serves some protective function. Behavioural and pathological signs of cadmium poisoning such as erratic swimming and blood clotting in alevins were observed.