Immunohistochemical localization of a gastrin-like peptide in the brain of an amphibian,Xenopus laevis Daud.

Summary The indirect immunofluorescence technique was used to demonstrate a substance reacting with gastrin antisera in the brain of Xenopus laevis.Immunoreactive material was found in two sites: (1) In the caudal hypothalamus more precisely in the nucleus infundibularis ventralis, (NIV) of the pars ventralis of the tuber cinereum, (PVTC). The fluorescent axons of the reactive parikarya of the NIV give rise to two symmetrical tracts which run rostro-ventrally and join, in the infundibular floor, the preoptico-hypophysial tract, where they form an uneven median tract coursing caudally and running along the medio-tuberal area before entering the external zone of the median eminence. (2) In the anterior preoptic area (APOA), where numerous nerve fibers and endings form a dense network near the preoptic recess. The exact origin of these terminals has not yet been determined.Control of immunohistochemical specificity shows that the labeling by gastrin antisera is suppressed by gastrin (2–17), but also by cholecystokinin (CCK) and pentagastrin (Peptavlon). These results indicate that the immunoreactive substance revealed belongs to the gastrin group and has an antigenic determinant composed of the amino acid sequence or a portion thereof common to gastrin, CCK and Peptavlon (Trp-Met-Asp-Phe).It should be emphasized that, in the brain of Xenopus laevis, both gastrinimmunoreactive sites correspond to the sites of uptake of steroid hormones (Kelley et al., 1975; Morrell et al., 1975).Supported by the D.G.R.S.T., Contrat n 77.7.0648     

Proopiomelanocortin (POMC)-related peptides in the brain of the rainbow trout, Salmo gairdneri

We have investigated the presence of ACTH, -MSH and β-endorphin, three peptides which derive from the multifunctional precursor protein proopiomelanocortin (POMC) in the brain of the rainbow trout Salmo gairdneri. Using both the indirect immunofluorescence and peroxidase-antiperoxidase techniques, a discrete group of positive cells was identified in the hypothalamus, within the anterior part of the nucleus lateralis tuberis. -MSH-containing neurons represented the most abundant immunoreactive subpopulation. Coexistence of -MSH, ACTH and β-endorphin was observed in the lateral part of the nucleus. ACTH- and β-endorphin-containing cells were mainly distributed in the rostral and caudal regions of the nucleus. In the medial portion of the nucleus lateralis tuberis, numerous cells were only stained for -MSH. Moderate to dense plexuses of immunoreactive fibers were observed in the ventral thalamus and the floor of the hypothalamus. Some of these fibers projected towards the pituitary. The concentrations of ACTH, -MSH and β-endorphin-like immunoreactivities were measured in microdissected brain regions by means of specific radioimmunoassays. Diencephalon, mesencephalon and medulla oblongata extracts gave dilution curves which were parallel to standard curves. The highest concentrations of POMC-derived peptides were found in the diencephalon (-MSH: 4.28±0.43 ng/mg prot.; ACTH: 1.08±0.09 ng/mg prot.; β-endorphin: 1.02±0.1 ng/mg prot.), while lower concentrations were detected in the mesencephalon, medulla oblongata and telencephalon. The present results demonstrate that various peptides derived from POMC coexist within the same cell bodies of the fish hypothalamus. Taken together, these data suggest that expression and processing of POMC in the fish brain is similar to that occurring in pituitary melanotrophs.     

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.     

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.     

Gonadotropic cells in the Atlantic salmon,Salmo salar

Summary The gonadotropin-producing cells (GTH-cells) in the Atlantic salmon were studied light and electron microscopically before, during and after spawning, and after injections of luteinizing hormone-releasing hormone (LH-RH). The double immunofluorescent technique was applied using rabbit anti-carp GTH as the first antibody. Numerous immunofluorescent cells were observed throughout the pars distalis, but very few in the pars intermedia. These cells are basophilic and PAS-positive, and ultrastructurally classified as globular gonadotropes. Only one gonadotropic cell type could be identified; its size, morphology and fine structure vary considerably. In the same specimen the GTH-cells can be predominantly globular or vesicular in appearance, depending on the reproductive phase of the fish. At spawning and after LH-RH injection, many GTH-cells reach a vacuolar stage; the content of the vacuoles is not immunofluorescent. Another cell type, which resembles GTH-cells in semithin sections, did not show gonadotropic properties; its nature and functional significance are unknown. In addition, the present study revealed an increase in the synthetic and exocytotic activity of prolactin cells after LH-RH injections. It is suggested that LH-RH mediates this effect via LH and eventually via estradiol.The authors are indebted to Dr. N. Johansson and superintendent N. Steffner for generously providing facilities at their institutes in Älvkarleby. They also wish to thank Mrs. Y. Lilliemarck and Mrs. V. Lundin for their skillful technical assistance. The grants made available by the Swedish Natural Science Research Council (No. 2124-037) are also gratefully acknowledged. Thanks are also due to Mrs. S.M. McNab for linguistic assistance     

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.     

Structure and function of the paraphysis cerebri in the rainbow trout,Salmo gairdneri Richardson

Summary The paraphysis cerebri of adult Salmo gairdneri is represented by a differentiated part of the pars impar telencephali of the telencephalic roof. It consists of a vaulted epithelial sheet, which displays only a few rostral evaginations and separates the cerebrospinal fluid (CSF) from the meningeal interstitial fluid. The fenestrated, sinusoidal portal system surrounding the paraphyseal epithelium appears to be part of a complex vascular bed of the dorsal telencephalic and diencephalic area. Myelinated and unmyelinated nerve fibers observed in the vicinity of the paraphyseal epithelium fail to make synaptic contact with paraphyseal cells. The single-layered epithelium is composed of characteristic, rather small, optically dense, cuboidal and cylindrical cells, apically mutually attached by junctional complexes including zonulae occludentes.These paraphyseal cells execute a high energetic and a moderate synthetic metabolism as indicated by ultrastructural, cytochemical and enzyme-cytochemical observations. Morphological evidence is presented for a multiple function of these cells in the regulation of the CSF: 1) water and solute elaboration into the ventricular system, 2) restricted uptake of high molecular weight organic substances from the CSF, 3) restricted uptake of low molecular weight substances from the CSF, but apparently not of GABA and of biogenic amines, 4) the formation and pinching-off of blebs as expression of a physiological mechanism not yet elucidated. The possible relationship between the level of development of the paraphysis cerebri and the sensitivity of animals to hydro-mineral metabolism is discussed.     

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.     

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.     

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.     

Polyploidy induced by hydrostatic pressure in rainbow trout, Salmo gairdneri Richardson

Triploid rainbow trout were produced by hydrostatic pressure applied to eggs 40 min after fertilization. Treatment for 10 min with or without exposure to 2% ether produced high hatching rates. Nuclear measurements from serial section of 40-day-old fry and from blood smears of 5-month-old juveniles showed that the proportion of triploid individuals was 80–90%. Ether treatment alone did not induce triploidy. Attempts to produce tetraploids by hydrostatic pressure treatment of eggs at 8 h after fertilization failed. Parallel results were also obtained with heat shock.     

Microvasculature of the pineal organ of the rainbow trout (Salmo gairdneri)

Summary The angioarchitecture of the pineal organ of the rainbow trout (Salmo gairdneri) was investigated by means of the corrosion-cast preparation method and scanning electron microscopy. Two main arteries (aa. epiphyseales) supply the pineal parenchyma. They emerge from the aa. cerebri anteriores and run in the fissure between the prosencephalon and the mesencephalon. After entering the pineal stalk, the aa. epiphyseales branch off into several arterioles, most of which extend to the pineal end-vesicle where they give rise to a lobular, bilaterally symmetric capillary network. Capillaries establishing the main portion of the pineal vessels appear widened in comparison to those supplying other portions of the brain and resemble capillaries in other endocrine organs. In Salmo gairdneri, no specialized system of portal vessels appears to exist between the pineal organ and other portions of the brain.     

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.     

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.     

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.     

Diploid gynogenesis induced by hydrostatic pressure in rainbow trout, Salmo gairdneri Richardson

Diploid gynogenetic fry of rainbow trout, Salmo gairdneri , were produced by treating eggs activated by UV-irradiated sperm with hydrostatic pressure. Treatments for l0 min at 40 min after activation with 55 000 kPa (8000 psi) pressure resulted in high hatching rates and yields of heterozygous diploid gynogenomes of up to 81%. Attempts to produce homo-zygous diploid gynogenomes by the suppression of the first mitotic division ofthe eggs failed. Ether treatment alone did not induce diploid gynogenesis.