Selenium-induced autometallographic demonstration of endogenous zinc in organs of the rainbow trout, Salmo gairdneri

Autometallographic (AMG) silver enhancement of endogenous zinc was studied in seven organs of the rainbow trout Salmo gairdneri. Groups of trout were injected intraperitoneally with sodium selenite in doses ranging from 0.08 to 25 ppm, administered 1 h before being killed. The concentration of selenium obtained by each organ was determined by gamma-spectrometry, and compared with the autometallographic deposition of silver grains. The relative accumulation of selenium in the organs was: liver greater than spleen greater than kidney greater than intestine greater than gills greater than brain greater than muscle. In the fish labelled with 10 and 25 ppm Se, AMG-deposits were found (1) within lysosomes of liver cells, (2) within the granules and on the nuclear membrane of melanophores in the spleen, (3) on the microvilli and in the apical cytoplasm of renal proximal tubular cells, (4) within the granules and along the plasma membrane of intestinal eosinophilic granule cells, and in the apical portion of the intestinal epithelium, and (5) in the gills, within granule cells and on the surface of the ionocytes. In the trouts injected with 5 ppm Se, silver grains were still observed in the liver, the intestine, and the gills, whereas, no such grains were found in preparations from fish having received 1 ppm Se. The use of selenium for the histochemical demonstration of endogenous zinc versus exogenous metals is discussed. Also, consideration is given to the question of which part of the total tissue zinc that is histochemically reactive.     

Uptake and accumulation of zinc in juvenile rainbow trout,Salmo gairdneri

Synopsis The toxicity of zinc to rainbow trout was determined and the 72 h median lethal concentration was found to be 2.00 mg l^–1 in freshwater, hardness 7.50 mg l^–1 as calcium. An insignificant increase in zinc concentration of internal tissues occurred in fish exposed to 1.52 mg l^–1 in freshwater for 72 h. However, there was a significant uptake of zinc by gills and the body surface. Fish exposed to 10 mg l^–1 zinc for 72 h in two-thirds sea water showed significant zinc uptake by liver, rectum and muscle, when compared to control fish. Drinking rate decreased from 1.43 to 0.26 ml kg^–1 h^–1 when zinc sulphate was added to freshwater. Trout adapted to two-thirds sea water showed no decrease in drinking, about 7 ml kg^–1 h^–1 when zinc was added to the water.     

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.     

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.     

Cytochemical demonstration of acid phosphatase activity in the pineal organ of the rainbow trout,Salmo gairdneri

Summary Activity of acid phosphatase (ACP) was investigated cytochemically in the pineal organ of the rainbow trout, Salmo gairdneri. Intense reaction product for ACP activity was observed (1) in lysosomes varying in size and shape and (2) in endoplasmic reticulum associated with the Golgi complex of (i) the pineal photoreceptor and supporting cells, (ii) vascular endothelial cells, and (iii) macrophages inhabiting pineal lumen, parenchymal epithelium and perivascular spaces. This localization of ACP is discussed with particular reference to the capacity for lysosomal digestion in a pineal organ combining photoreceptive and secretory functions, and lacking a blood-brain barrier, as holds true for the pineal of the rainbow trout. Taking advantage of its capacity for endocytotic uptake and lysosomal digestion, the pineal organ of the rainbow trout may serve as a barrier between the blood circulation and the cerebrospinal-fluid compartment. Furthermore, the macrophages may be considered as an essential component in pineal function of fish.Fellow of the Alexander von Humboldt Foundation.Fellow of the Alexander von Humboldt Foundation.     

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 autoradiographic localization of zinc within the pancreatic islets of the rainbow trout,Salmo gairdneri

Summary This experiment reports on the localization of zinc within the pancreatic islets of Salmo gairdneri. Individual fish were injected with ⁶⁵Zn and the distribution of the isotope within the islets was determined by autoradiography. Insulin cells were found to accumulate approximately twice as much zinc per unit area as the rest of the islet tissue. It is presumed that this zinc is involved with the crystallization and storage of insulin within the insulin cells. Various histological methods were investigated to ascertain the procedure which best precipitated zinc and at the same time avoided excess leaching.     

The autoradiographic localization of zinc within the pancreatic islets of the rainbow trout, Salmo gairdneri

This experiment reports on the localization of zinc within the pancreatic islets of Salmo gairdneri. Individual fish were injected with 65Zn and the distribution of the isotope within the islets was determined by autoradiography. Insulin cells were found to accumulate approximately twice as much zinc per unit area as the rest of the islet tissue. It is presumed that this zinc is involved with the crystallization and storage of insulin within the insulin cells. Various histological methods were investigated to ascertain the procedure which best precipitated zinc and at the same time avoided excess leaching.     

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.     

Agglutination of spermatozoa by autoantibodies in the rainbow trout, Salmo gairdneri

Agglutinating antibodies to self spermatozoa were induced in mature male rainbow trout, whether immunised with autologous spermatozoa or allogeneic testis. Both sperm heads and flagellae appeared to be autoantigenic. These are the first results which show that fish can produce autoantibodies against spermatozoa. This is an important step towards the control of reproduction in fish using vaccines.     

Glomerular binding of angiotensin II in the rainbow trout,Salmo gairdneri

Summary Isolated glomeruli of the rainbow trout have been exposed in vitro to¹²⁵I-angiotensin II (0.88 × 10^–9 M) and binding sites located by light-microscopic autoradiography. These studies provide evidence of specific binding of angiotensin II by glomeruli. Binding was significantly inhibited by excess (10^–5 M) unlabelled angiotensin II, but a high degree of non-specific binding also occurred. The mammalian competitive antagonist, saralasin (3 × 10^–7 M) did not influence¹²⁵I-angiotensin II binding to fish glomeruli. Intense binding of¹²⁵I-angiotensin II was noted at the vascular pole of some glomeruli.     

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.     

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.     

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.