Localization by immunofluorescence of a gastrin-like substance in the brain of the rainbow trout,Salmo gairdneri

Summary With the aid of an indirect immunofluorescence technique neurones containing a gastrin-like substance were identified in the brain of Salmo gairdneri. The perikarya of these neurones appear to be located along the periventricular part of the nucleus lateralis tuberis between the hypophysial stalk and the most rostral tip of the saccus vasculosus. The fibres of these perikarya run rostrally toward the hypophysis, where they can be followed in the protrusions of the neurohypophysis into the proximal pars distalis. Here the bundle of immunoreactive fibres divides into numerous smaller bundles and into single fibres. Immunohistochemical specificity tests have shown this immunoreactive substance to belong to the gastrin group, sharing an antigenic determinant with cholecystokinin (CCK) and pentagastrin (common aminoacid sequence Trp-Met-Asp-Phe). A possible function of these gastrin (or CCK)-containing neurones in the rainbow trout is discussed.     

DDT induced ethological changes in estuarine fish

Synopsis Experiments were carried out to study the feeding rates of the predator fish Therapon jarbua (Forsk) on mullet juveniles, before and after treatment with DDT. Mullet juveniles treated with a subacute concentration, were refused by the control predators, whereas predators treated with a subacute concentration consumed more mullet juveniles. In the present study crescent perch T. jarbua were exposed to subacute and acute concentrations of DDT, and their behaviour was compared with that of the control predators. There were changes in oriented behaviour and co-ordinated movements, and in feeding, aggression and comfort behaviour of the fish. Inflammation in the gills, and caudal fin serration, were noticed in treated fishes. The findings presented here throw light on fundamental pathways by which pollutants interact with the behaviour of fishes.     

Structural studies of nerve terminals containing melanin-concentrating hormone in the eel,Anguilla anguilla

Summary Eels were adapted to black- or white-coloured backgrounds and the pituitary glands were prepared for light and electron microscopy. Immunocytochemical staining was used to study the distribution of the neurohypophysial melanin-concentrating hormone in the neurointermediate lobe. The hormone was located in small, elliptical, electron-opaque neurosecretory granules, measuring approximately 120×90 nm. The neurones terminated on blood vessels in the centre of the neurohypophysis and on the basement membrane separating neural and intermediate lobe tissues. The results of both light and electron immunocytochemistry and of radioimmunoassay are consistent with a higher rate of hormone release from eels adapted to white backgrounds than from those adapted to black backgrounds. In addition to this, when fish that had been adapted to white tanks were transferred to black tanks, there was an accumulation of irMCH in the gland and an increased numerical density of secretory granules at nerve terminals. These results reinforce the proposal that MCH is released during adaptation to a white background, to cause melanin concentration and to inhibit MSH release, and that its release is halted in black-adapted fish.     

Distribution of salmon gonadotrophin releasing-hormone in the brain and pituitary of the sea bass (Dicentrarchus labrax)

Summary The distribution of salmon gonadotrophin-releasing hormone (sGnRH) was studied in the brain and pituitary of two-year-old immature sea bass (Dicentrarchus labrax) by means of an enzymoimmunoassay (EIA) for sGnRH and immunocytochemistry. The EIA for sGnRH is a competitive assay using a tracer made of sGnRH coupled to acetylcholinesterase from an electric eel. The separation of free and bound tracer is achieved by coating the plates with mouse anti-rabbit IgG monoclonal antibodies. Displacement curves generated by sGnRH and extracts from pituitary and different brain regions showed a good parallelism allowing the assay to be used for sGnRH measurements in this species. Although all parts of the brain contained measurable levels of sGnRH, the highest concentrations were found in the pituitary, the olfactory bulbs and the telencephalon. These data were confirmed by immunocytochemistry. Cell bodies were found in the olfactory bulbs, ventral telencephalon, preoptic region and mediobasal hypothalamus. Immunoreactive fibers could be observed in all parts of the brain including the optic tectum, the cerebellum (corpus and valvula), the vagal lobe, the medulla oblongata and the rostral spinal cord. In most cases, these fibers do not form well defined bundles; however, there was clearly a continuum of immunoreactive fibers, extending from the olfactory bulbs to the pituitary, and along which all the cell bodies described above were located. In the ventral telencephalon and the preoptic region, clear pictures of varicose positive fibers contacting immunoreactive perikarya could be observed. These data indicate that sGnRH is most likely an endogenous peptide in the brain of the sea bass, although the presence of other forms of GnRH cannot be excluded at this point. This study also demonstrates that the general organization of the GnRH systems in the sea bass is highly similar to what has been described in most freshwater teleost species, and provides basis for further studies on the neuroendocrine control of gonadotrophin release in this commercially important species.     

Phosphoglucose isomerase isozymes in teleostean fishes from the Arabian Gulf

Two phosphoglucose isomerases (PGI) with different electrophoretic mobilities have been found in all groups of teleostean fishes studied, with the exception of the Clupeomorpha. PGI proved to be a good taxonomic criterion to differentiate members of the Nemipteridae, Sciaenidae, Platycephalidae and Stromateidae from the other teleost families.     

Analysis of two chloride requirements for sodium-dependent amino acid and glucose transport by intestinal brush-border membrane vesicles of fish

Intestinal brush border vesicles of a Mediterranean sea fish (Dicentrarchus labrax) were prepared using the Ca²⁺-sedimentation method. The transport of glucose, glycine and 2-aminoisobutyric acid is energized by an Na⁺ gradient ($\text{out > in}$). In addition, amino acid uptake requires Cl^? in the extravesicular medium (2-aminoisobutyric acid more than glycine). This Na⁺- and Cl^?-dependent uptake is electrogenic, since it can be stimulated by negative charges inside the vesicles. The specific Cl^? requirement of glycine and 2-aminoisobutyric acid transport is markedly influenced by pH, a change from 6.5 to 8.4 reducing the role played by Cl^?. In the presence of Cl^?, the $\text{K}{}_{\mathrm{<mtext>m</mtext>}}$ of 2-aminoisobutyric acid uptake is reduced and its $\text{V}{}_{\mathrm{<mtext>max</mtext>}}$ is enhanced. Cl^? affects also a non-saturable Na⁺-dependent component of this amino acid uptake. Amino acid transport is also increased by intravesicular Cl^? (2-aminoisobutyric acid less than glycine). This effect is more concerned with glucose uptake, which can be then multiplied by 2.3. A concentration gradient ($\text{in > out}$) as well as the presence of Na⁺ in the incubation medium seems to enter into this requirement. This intravesicular Cl^? effect is not influenced by pH between 6.5 and 8.4.     

Zur Charakterisierung neuronaler und gliöser Elemente im Epithel des Saccus vasculosus von Knochenfischen

Zusammenfassung Das Epithel des Saccus vasculosus des Flußbarsches Perca fluviatilis besteht aus Krönchenzellen, bipolaren Liquorkontaktneuronen (Zahlenverhältnis etwa 41) und Stützzellen. Im Bereich des Saccus kommen Macrophagen vor. Die Krönchenzellen wurden unter verschiedenen Fixierungsbedingungen untersucht. Die Globuli enthalten schlauchförmige Zisternen, die nicht mit den Zisternen des Zellapex in Verbindung stehen. Im Zytoplasma des Zellapex und der Globuli wird bei langdauernder OsO₄-Imprägnation Osmium gebunden. Die Krönchenzellen werden basal von Ausläufern der Stützzellen unterlagert. Sie werden nicht innerviert und entsenden keine Axone.Die bipolaren Neurone sind durch einen im Liquor endigenden Dendriten und ein Axon gekennzeichnet, das in eines der Faserbündel des Epithels eintritt. Der Dendrit trägt 1 bis 2 Zilien. Die Zelle ist reich an Vesikeln und kann am Perikaryon wie an den Ausläufern Synapsen tragen. Im extrazellulären Raum um die Neurone und in vesikulären Strukturen des Apex wird Acetylcholinesterase nachgewiesen.Der Nervus sacci vasculosi dürfte nur afferente Axone von Liquorkontaktneuronen und efferente Fasern, die diese innervieren, enthalten.

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Neuronal and glial cell elements within the epithelium of the Saccus vasculosus in teleosts

Summary The epithelium of the Saccus vasculosus of Perca fluviatilis consists of coronet cells and bipolar liquor contact neurones in a 41 ratio, and supporting cells. The organ also contains macrophages.The coronet cells have been studied after different kinds of fixation. The globules of these cells contain tubelike cisternae, which do not connect with cisternae in the cell's apical protrusion. The cytoplasm of the apical protrusion and to a greater extent of the globules, is stained by longlasting OsO₄-impregnation. The coronet cells have no direct contact with the basement membrane of the organ. They are neither innervated nor have axons.The dendrites of the bipolar nerve cells end with a bulbous structure protruding into the cerebrospinal fluid. The dendrites contain vesicles and each bears 1–2 cilia. The axons join the fiber bundles of the epithelium. There are synaptic contacts on the surface of the neurons and their processes. In some vesicular structures within the apices and more conspicuously within the extracellular space around these cells indications of acetylcholinesterase activity are found.It appears that the nervus sacci vasculosi contains only afferent axons of the bipolar liquor-contact neurons and efferent fibres which form synaptic contacts with these neurons.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Ionic Requirements for Catecholamine Aggregating Actions on the Teleost Poecilia reticulata Melanophores

Norepinephrine (NE) and phenylephrine (Phe) were employed to study the ionic requirements for alpha adrenoceptor activation in the teleost Poecilia reticulata melanophores. As expected the beta adrenoceptor blocker, propranolol, increased the sensitivity of the preparation to NE (5.8 times), and was therefore employed in all the experimental procedures. Neither cocaine (a neuronal uptake blocker) nor dexamethasone (an extraneuronal uptake blocker) enhanced the sensitivity of the preparation to NE, suggesting that these inactivating mechanisms would not play a role in P. reticulata pigmentary system. However, in the absence of calcium, the dose-response curve (DRC) to NE was displaced to the left about 3.5 times, whereas the DRC to Phe was not affected. These results indicate that a neuronal uptake is active, but was not demonstrated by the classical pharmacological tools, probably due to an assymmetric display of the nervous endings. The DRC to NE was rightward displaced (14.1 times) in the presence of the calcium channel blocker Verapamil, whereas the DRC to Phe was not affected. These data suggest that P. reticulata melanophores possess a mixed population of alpha₁ and alpha₂ adrenoceptors, the activation of the latter eliciting an extracellular calcium influx. In sodium-free saline, the DRC to NE was rightward shifted (6.6 times) and the response to Phe was impaired in such a way that the maximal response was not achieved. The DRC to both NE and Phe were rightward displaced (7.9 and 2.7 times respectively) in the presence of the sodium channel blocker tetrodotoxin (TTX) 10^?7M. In potassium-free saline, the melanophore sensitivity to Phe was increased, whereas the responses to NE were not affected, suggesting a differential sensitivity of the two alpha adrenoceptor subtypes to the resulting membrane hyperpolarization. Based on the literature and on our data we propose that P. reticulata melanophores possess a mixed population of alpha₁ and alpha₂ receptors. The activation of both subtypes of alpha adrenoceptors elicits a Na⁺ ion influx through TTX-sensitive sodium channels. The stimulation of alpha₂ adrenoceptors also requires an extracellular calcium influx, through the opening of slow calcium channels.     

Rhythmic melatonin secretion in different teleost species: an in vitro study

The rhythmic production of melatonin is governed by intrapineal oscillators in all fish species so far investigated except the rainbow trout. To determine whether the latter represents an exception among fish, we measured in vitro melatonin secretion in pineal organs of nine wild freshwater and six marine teleost species cultured at constant temperature and under different photic conditions. The results demonstrate that pineal organs of all species maintain a rhythmic secretion of melatonin under light:dark cycles and complete darkness, and strongly suggest that most fish possess endogenous intrapineal oscillators driving the rhythm of melatonin production, with the exception of the rainbow trout.Abbreviations LD light:dark - DD dark:dark - NAT N-acetyltransferase - RIA radioimmunoassay     

Spermatogenesis in the black porgy,Acanthopagrus schlegeli (teleostei: Perciformes: Sparidae)

The ultrastructure of spermatogenesis and of the spermatozoon of Acanthopagrus schlegeli (Sparidae) are described. The testis is of the unrestricted type. Germ cells are surrounded by cyst cells. Spermiogenesis involves conspicuous modifications such as intracellular movements (diplosome and mitochondria migration, nuclear rotation, and depression) and structural changes (chromatin condensation, shape of mitochondria, and loss of cytoplasm). The mature spermatozoon has a spherical nucleus with a deep, axial nuclear fossa, and an unusual notch, shaped like a bow tie. The short midpiece contains four spherical mitochondria and encircles the basal body of the flagellum. It is concluded that the A. schlegeli spermatozoon is of a primitive type, but that it is characterized by a unique feature which may provide a useful systematic character. © 1993 Wiley-Liss, Inc.