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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Journal of fish biology 47 (1995), S. 0 
    ISSN: 1095-8649
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A study of the annual ovarian cycle of the flounder, Pleuronectes flesus, was made, describing the different stages of germ cell maturation and the different phases of ovarian development. The development of oocytes in the flounder is a group synchronous process with two distinct generations of primary growth phase oocytes. One group becomes vitellogenic at the end of the summer while the other group does not surpass the primary growth phase. Oogonia, as well as seven stages of oocyte development and post-ovulatory follicles were distinguished. The occurrence of these oocyte stages was used to define the different phases of ovarian development. Previtellogenesis and vitellogenesis were each divided into two subphases: the perinucleolus and cortical alveoli phase, and early and advanced vitellogenesis, respectively. These phases were followed by final maturation, ovulation, spawning and a post-spawning phase. The GSI increased rapidly during vitellogenesis, to a maximum value of about 35 just before spawning, at the end of the winter, when the animals had migrated to the North Sea spawning grounds. After spawning the GSI dropped to 1.5. Fish that had lived in a polluted environment exhibited premature vitellogenesis, resulting in a high number of oocytes in yolk granule stage. This study on the effects of pollution indicates the importance of the description of the normal annual cycle of the ovary as a control in toxicological studies.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Naturwissenschaften 55 (1968), S. 393-394 
    ISSN: 1432-1904
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Histochemistry and cell biology 97 (1992), S. 133-139 
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pituitaries of the African catfish, Clarias gariepinus, were prefixed in aldehyde fixatives, frozen in liquid propane and submitted to a cryosubstitution procedure. Ultrathin sections of the Lowicryl HM20-embedded tissue were treated with primary antisera raised in rabbits to gonadotropin releasing hormone (GnRH), vasopressin or gamma amino butyric acid (GABA) respectively. Binding of the primary antisera was visualized with goat anti-rabbit (GAR) labeled with gold. The general morphology of the tissue components in the cryosubstituted pituitaries matches with that obtained after routine embedding procedures. In addition, a strong labeling intensity of the neuropeptides/neurotransmitters investigated in the present study was demonstrated. Due to these qualities cryosubstitution provides optimal conditions for studying co-localization of neurosecretory products, using double-immunostaining procedures. In the pars distalis of the catfish pituitary several types of hypothalamus-derived nerve fibers are present between or synapting on the secretory cells. It is demonstrated that the two known catfish GnRHs are co-localized in the same nerve fiber and within these nerve fibers even co-exist in the same neurosecretory granules. GABA and vasopressin-immunolabeling each occurred in different nerve fibers. The present data demonstrate that cryosubstitution and low temperature-embedding results in an excellent morphological preservation compared to ultracryotomy and a better preserved immunoreactivity of small antigenic molecules in comparison to conventional fixation and embedding techniques.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-5168
    Keywords: Clarias gariepinus ; African catfish ; teleost fish ; puberty ; spermatogenesis ; steroidogenesis ; testis ; interrenal tissue ; androgen blood levels
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In fish, sex steroids initiate and/or accelerate the maturation of the brain-pituitary-gonad axis. In order to obtain information on the steroid milieu during the pubertal development of male African catfish, we have monitored the conversion of [3H]-pregnenolone and [3H]-androstenedione by testis and [3H]-pregnenolone by interrenal tissue fragmentsin vitro. Pubertal development occurs between two and six months of age. Testicular development proceeds through four main stages that are characterised histologically by the presence of spermatogonia (stage I), spermatogonia and spermatocytes (stage II), spermatogonia, spermatocytes and spermatids (stage III), and all germ cells including spermatozoa (stage IV). 11β-Hydroxyandrostenedione and cortisol were the main products of testes and interrenal tissue, respectively, in all stages of the pubertal development; a change in the steroidogenic pattern was not observed during this period. The interrenal tissue displayed no significant conversion of [3H]-pregnenolone to 11-oxygenated androgens. Blood plasma was analyzed for the presence of five androgens; testosterone, 11β-hydroxytestosterone, 11β-hydroxyandrostenedione, androstenetrione, and 11-ketotestosterone. 11-Ketotestosterone was the quantitatively dominating androgen in the circulation at all stages of development, which was more pronounced in stages III and IV. The obvious differences between thein vitro andin vivo results, namely 11β-hydroxyandrostenedione being the main testicular productvs. 11-ketotestosterone dominating in the blood, may indicate that 11β-hydroxyandrostenedione is converted to 11-ketotestosterone at extratesticular sites.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1573-5168
    Keywords: 11-ketotestosterone ; gonadotropin releasing-hormone ; gonadotropin secretion ; steroid feedback ; male African catfish
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Description / Table of Contents: Résumé La 11-ketotestosterone (OT) est un androgène caractéristique des poissons téléostéens mâles mais les informations sur son implication dans le rétrocontrôle sur la sécrétion de l'hormone gonadotrope II (GtH II) sont très contreversées. Les effets de OT sur la sécrétion de GtH II induite par la gonadolibérine (GnRH) ont donc été étudiés chez le poisson chat africain (Clarias gariepinus) dans le présent papier. Des expériences ont été menées in vivo sur des poissons intacts et castrés. Les teneurs plasmatiques en OT ont été augmentées grâce à l'implantation de capsules en silastic contenant de la 11-ketoandrostenedione (OA) qui est transformée en OT chez les poissons intacts et castrés. Quand les mâles intacts reçoivent des implants témoins ou contenants de l'OA, un traitement avec un analogue de la gonadolibérine de saumon (Des-Gly10-DArg6-sGnRH-Net; 0.2 μg de sGnRHa/kg de poids vif) entraîne une élévation des teneurs plasmatiques de GtH II dans les deux groupes d'animaux. Cependant, les niveaux plasmatiques sont 2 fois plus élevés chez les poissons témoins que chez ceux implantés avec de l'OA. Quand les poissons castrés reçoivent des implants témoins ou contenant de l'OA, le traitement au sGnRHa induit une augmentation de la GtH plasmatique plus élevées que celle observée chez les poissons “fantômes”. Cependant, il n'y a pas de différence entre les poissons castrés témoins et implantés avec OA bien que l'implantation avec OA entraîne une restauration des niveaux plasmatiques en OT. Ceci suggère que la supplémentation en OT ne suffit pas à renverser les effets induits par la castration. Des expériences in vitro ont été menées en utilisant des fragments hypophysaires maintenus dans un système de culture statique. Les fragments hypophysaires restent sensibles au sGnRHA (5 × 10−9M) pendant 4 jours. Une préincubation pendant 24 heures avec 25 ng de OT par ml de milieu (80 nM) abolit complètement l'effet stimulateur du sGnRHa sur la sécrétion de GtH II. L'OT tritiée n'était pas métabolisée pendant 8 heures d'incubation. En conclusion, chez le poisson chat african mâle, la 11-ketotestostèrone, androgène circulant prédominant et non aromatisable participe, au moins en partie par une action directe au niveau hypophysaire, à la régulation d'un rétrocontrôle négatif sur la sécrétion de GtH-II induite par le GnRH.
    Notes: Abstract 11-ketotestosterone (OT) is a typical androgen of male teleost fish, but information on the question if it is involved in the feedback regulation of pituitary gonadotropin II (GTH-II) secretion is controversial. We have therefore studied the effects of OT on gonadotropin releasing-hormone (GnRH) stimulated GTH-II secretion in male African catfish Clarias gariepinus). In vivo experiments were carried out with intact and castrated fish. OT plasma levels were increased by implantation of silastic capsules containing 11-ketoandrostenedione (OA) which is converted to OT in both intact and castrated fish. When intact males received OA- or blank-capsules, treatment with salmon gonadotropin releasing-hormone analogue (Des-Gly10-D-Arg6-sGnRH-NEt; 0.2 μg sGnRHa/kg body weight) elevated the plasma GTH-11 levels in both groups. However, the levels were about 2 times higher in blank- than in OA-implanted fish. When castrated fish received either blank-or OA-capsules, sGnRHa treatment led to plasma GTH levels significantly higher than in sham-operated fish. However, there was no difference between the blank- or OA-implanted castrates, though OA implantation led to a restoration of OT plasma levels. This suggests that replacement ofOT is insufficient to reverse castration-induced effects. In vitro experiments were carried out with pituitary tissue fragments using a static culture system. The tissue remained sensitive to sGnRHa (5 × 10−9M) for 4 days after the beginning of incubation. Preincubation of pituitary tissue for 24 hours with 25 ng OT/ml medium (80 nM) completely abolished the stimulatory effect of sGnRHa on GTH-II secretion. Tritiated OT was not metabolized by pituitary tissue during 6 hours of incubation. We conclude that 11-ketotestosterone, a quantitatively prominent and non-aromatizeable circulating androgen participates, at least in part by direct action on the pituitary, in the negative feedback regulation of GnRH-stimulated GTH-II secretion in male African catfish.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1573-5168
    Keywords: Carassius auratus ; Clarias gariepinus ; reproduction ; gonadotropin ; luteinining hormone releasing hormone (LHRH) ; receptors ; steroid ; castration ; sexual cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The goldfish pituitary contains two classes of gonadotropin-releasing hormone (GnRH) binding sites, a high affinity/low capacity site and a low affinity/high capacity site (Habibiet al. 1987a), whereas the catfish pituitary contains a single class of high affinity GnRH binding sites (De Leeuwet al. 1988a). Seasonal variations in pituitary GnRH receptor binding parameters, and the effect of castration on pituitary GnRH receptor binding were investigated in goldfish and catfish, respectively. In goldfish, GnRH receptors undergo seasonal variation with the highest pituitary content of both high and low affinity sites occurring during the late stages of gonadal recrudescence. The observed changes in pituitary GnRH receptor content correlate closely with responsiveness to a GnRH agonistin vivo in terms of serum gonadotropin (GTH) levels. In catfish, castration results in a two-fold increase in pituitary GnRH receptor content, which can be reversed by concomitant treatment with androstenedione, but not by the non-aromatizable androgen 11β-hydroxyandrostenedione; changes observed in GnRH receptor content correlate with variations in serum GTH levels and responsiveness to a GnRH agonist. In summary, the present study provides a clear evidence for seasonal variation in pituitary GnRH receptor activity in goldfish, and demonstrates a gonadal feedback mechanism regulating GnRH receptor activity in the catfish pituitary.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1573-5168
    Keywords: catfish ; pituitary ; GnRH ; receptor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Receptors for gonadotropin-releasing hormone (GnRH) were characterized using a radioligand prepared from a superactive analog of salmon GnRH (sGnRH), D-Arg6-Pro9-sGnRH-NEt (sGnRHa). Binding of125I-sGnRHa to catfish pituitary membrane fractions reached equilibrium after 2 h incubation at 4°C. Displacement experiments with several GnRH analogs as well as other peptides, demonstrated the specificity of125I-sGnRHa binding. Specific binding was enhanced in the presence of the cation chelator ethylene bis (oxyethylenenitrilo) tetra-acetic acid (EGTA), indicating an inhibitory effect of cations on GnRH-receptor binding. The binding of125I-sGnRHa to pituitary membranes was found to be saturable at radioligand concentrations of 5 nM and above. A Scatchard analysis of the saturation data suggested the presence of a single class of high-affinity binding sites (Ka=0.901±0.06×109M−1, Bmax=1678±150 fmol/mg protein). A comparative study on125I-sGnRHa binding to pituitary membrane fractions of male and female catfish, indicated that there were no differences in binding affinity and binding capacity between both sexes. The results demonstrate the presence of specific, saturable GnRH receptors in the African catfish pituitary.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1573-5168
    Keywords: COMT ; gonadotropic cells ; dopamine ; catecholestrogens ; Clarias gariepinus ; teleosts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Isolated gonadotrops of the African catfish,Clarias gariepinus, were incubated with dopamine (DA) and/or catecholestrone and the activity of the enzyme catechol-O-methyltransferase (COMT) was determined by measuring the methylated products. From the apparent Km values for DA and catecholestrone of 0.4–1.3 μM and 17.9–25.2 μM respectively, it was concluded that catecholestrone is a better substrate for the enzyme COMT, compared to DA. Moreover, the methylation of DA is inhibited by comparatively low concentrations of catecholestrone.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1573-5168
    Keywords: gas chromatography-mass spectrometry ; teleosts ; testis ; steroids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract With the final aim of identifying the testicular steroids involved in the feedback mechanism of the hypothalamus-pituitary-gonad axis, steroid secretion by the testis of African catfish, Clarias gariepinus, was studied in vitro, by means of gas chromatography followed by mass spectrometry. Testicular fragments of sexually mature catfish raised in captivity were incubated in L-15 medium with and without catfish pituitary extract (cfPE). Without adding cfPE, 22 steroids could be identified, amongst which 11β-hydroxyandrostenedione, 11β-hydroxytestosterone, 11-ketotestosterone and 11-ketoandrostenedione were dominating. After incubation in the presence of cfPE, the concentrations of the four 11-oxygenated steroids were increased about 4-fold. The amounts of pregnane derivatives in the incubation medium showed the largest increases in the presence of cfPE. 5β-Pregnane-triol levels, for example, were 60-fold higher than in the medium from control incubations. The secretion of 5α- and 5β-androstanes was also stimulated by cfPE. The stimulation was not equal for all steroids, indicating that cfPE not only stimulates total steroidogenesis by increasing the availability of cholesterol, but also by influencing specific steroid converting enzyme activities.
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  • 10
    ISSN: 1573-5168
    Keywords: catfish ; pituitary ; ontogeny ; gonadotrops ; ultrastructure ; stereological analysis ; immunocytochemistry ; GTH subunits ; plasma androgen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Description / Table of Contents: Résumé A l'aide d'une étude d'immunocytochimie ultrastructurale, nous avons suivi chez le poisson chat africain mâle âgé de 2 à 6 mois le développement des cellules gonadotropes hypophysaires en relation avec celui des testicules. Pendant cette période, 5 prélèvements hypophysaires et testiculaires ont été effectués (groupe I–V) alors que les prélèvements sanguins n'ont pu être pratiqués que dans les groupes III–V. Le développement testiculaire a été divisé en 3 stades c'est à dire immature (spermatogonies seulement, groupe I), début de spermatogenèse (spermatogonies et spermatocytes, groupe II et III), spermatogenèse avancée (tous les stades de cellules germinales incluant les spermatozïdes, groupe IV et V). Les niveaux sanguins en 11-ketotestostèrone étaient bas excepté dans le dernier groupe. Des anticorps ont été fabriqués contre l'α,βGtH-II et les sousunités α et β de GtH-II de poisson chat. Dans la partie proximale distale de l'hypophyse du poisson immature, des cellules indifférenciées, des cellules somatotropes, des cellules putatives thyrotropes (pTSH) et gonadotropes (pGtH) ont été trouvées. Dans les deux derniers types cellulaires, des granules de sécrétion ont été marqués avec l'anti-αGtH mais pas avec l'antiβ-GtHII. Les cellules présumées à TSH et GtH ont été distinguées par la taille des granules de sécrétion. Pendant le début de la spermatogenèse, 2 classes de cellules gonadotropes putatives ont pu être distinguées. Un type a les mêmes caractéristiques immunocytochimiques et ultrastructurales que chez les immatures; les granules de sécrétion dans le second type cellulaire, qui était plus abondant, répondaient aussi immunopositivement à l'anti-βGtHII. Le volume moyen des granules de sécrétion de ces cellules à GtHII était 3 fois supérieur à celui des cellules putatives à GtH apparaissant tôt. De plus, les cellules à GtHII apparaissant tardivement contiennent de grandes inclusions, connues sous le nom de globules. Ces structures étaient marquées avec l'anti-αβGtHII et avec l'anti-βGtHII, mais pas avec l'anti-αGtH. Il a été supposé que les globules sont impliqués dans un stockage différentiel et/ou destruction des sous-unités de GthII. Pendant les stades plus avancés de la spermatogenèse les 2 typev gonadotropes pouvaient encore être distingués, mais les cellules putatives à GtH apparaissant tôt étaient rares. Les présentes observations permettent de supposer que les cellules à GtH apparaissant tôt peuvent être des cellules à GtHI. Cependant, la preuve définitive de leur identité dépend de la disponibilité d'anti-GtHI ou de cDNAs spécifiques de la GtHI.
    Notes: Abstract In an ultrastructural immunocytochemical study we investigated the development of the gonadotropic cells in the pituitary of two to six months old male African catfish in relation to testicular development. In this period, pituitary and testicular tissue samples were collected on five occasions (groups I–V). Blood samples could only be taken from the fish in groups III–V. The testicular development was divided in three stages i.e., immature (only spermatogonia, group I), early (spermatogonia and spermatocytes, groups II and III) and advanced (all germ cell stages including spermatozoa, groups IV and V) spermatogenesis. 11-Ketotestosterone blood levels were low, except for the last group. Antisera were raised against the complete catfish α,βGTH-II, as well as to the separate α- and β-subunits of catfish GTH-II. In the proximal pars distalis of immature fish, undifferentiated cells, somatotrops, putative thyrotrops (pTSH) and putative gonadotrops (pGTH) were found. In the two latter, secretory granules were labeled with anti-αGTH, but not with anti-βGTH-II. pTSH- and pGTH-cells were distinguished on the basis of the size of their secretory granules. During early spermatogenesis, two classes of putative gonadotrops could be distinguished. One type had the same immunocytochemical and ultrastructural characteristics as in immature fish; the secretory granules in the second cell type, which was more abundant, were also immunopositive for anti-βGTH-II. The mean volume of the secretory granules in these GTH-II cells was three times larger than that in the early appearing pGTH-cells. In addition, the later appearing GTH-II cells contained large inclusions, known as globules. These structures labeled with anti-αβGTH-II and with anti-βGTH-II, but not with anti-αGTH. It is assumed that the globules are involved in a differential storage and/or breakdown of the GTH-II subunits. During advanced spermatogenesis the two gonadotropic cell types could still be distinguished, but the early appearing pGTH-cell type was scarce. The present observations permit the conclusion that the early appearing cells may be GTH-I cells. However, definitive proof about their identity depends on the availability of antibodies or cDNA probes specific for GTH-I.
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