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  • 1
    ISSN: 1432-0878
    Keywords: Pituitary ; Teleost ; GTH-cells ; Innervation ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the male black molly, Poecilia latipinna, morphological and functional aspects of the gonadotropic (GTH-)cells have been studied at the ultrastructural level. The cells exclusively occupy the ventral and lateral areas of the meso-adenohypophysis. In the black molly there is evidence of the presence of only one type of gonadotropic cell. In the GTH-cells of most specimens, the rough endoplasmic reticulum is weakly developed. The secretory vesicles are characterized by cores with varying diameters; this variation was not observed in the secretory vesicles of the other types of pituitary cells, except in the TSH-cells. After applying a histochemical method for the demonstration of polysaccharides, small black deposits appear in the core of the secretory vesicles of the GTH and TSH-cells only; this indicates the glycoproteinaceous nature of the hormones produced in these cells. Male black mollies treated with methyl-testosterone have significantly smaller GTH-cells and a lesser number of secretory vesicles and mitochondria in these cells. GTH-cell activity in Poeciliinae may be thus influenced by androgens by means of a negative feed-back mechanism. The GTH-cells are innervated by both type A and type B neurosecretory fibres. There are indications that the type A fibres may originate from the pars lateralis cells of the nucleus lateralis tuberis; the origin of the type B fibres is uncertain.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0878
    Keywords: Gonadotropin Releasing Hormone ; Fore-brain ; Rana esculenta ; Immunofluorescence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using antibodies against mammalian LH-RH, the double antibody immunofluorescence technique has been applied to serial cross sections of the brains of adult Rana esculenta. Immunoreactive material was found in perikarya of an unpaired nucleus in front of the preoptic recess. The axons of these perikarya also contain fluorescing material. They form a single bundle which passes under the preoptic recess, than splits into two tracts, one on either side of the optic chiasm. The two tracts reunite just before entering the median eminence. The axons end near the capillaries in the outer zone of the median eminence. The possibility of two separate centres for the stimulation of gonadotropic activity in the brains of anurans is discussed.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0878
    Keywords: Gonadotrops ; Isolation ; Cell culture ; Gonadotropin secretion ; African catfish (Clarias lazera)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Dispersed pituitary cells from male African catfish, Clarias lazera, were fractionated in a density gradient of Percoll. Five fractions were isolated, consisting of about 6, 19, 39, 95 and 83% gonadotrops, respectively. The gonadotrops were identified by their ultrastructural characteristics, by immunocytochemistry, and by measuring their hormone content. After one day in culture, in each fraction the secretion of gonadotropin could be stimulated by a luteinizing hormone-releasing hormone analogue, indicating that the cells had retained their functional integrity. Since the regulatory mechanisms of different cell types from the pituitary have some similarity, purification of the gonadotrops provides a model to study the regulation of gonadotropin secretion.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-0878
    Keywords: Pituitary gland, pars anterior (distalis) ; Gonadotrops ; Thyrotrops ; Ultrastructure ; Immunolabeling ; Teleosts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pituitaries of the African catfish (Clarias lazera) were studied with immunocytochemical methods, at the light-microscopic and ultrastructural levels, for the characterization and localization of gonadotropic and thyrotropic cells. Two immunostaining procedures with the use of different markers were carried out: (i) with peroxidase-antiperoxidase, (ii) with protein A-gold. In routinely stained sections for light microscopy two types of basophils were identified in the proximal pars distalis: (1) large, round, purple cells, and (2) small, angular, light-blue cells. Both types were immunolabeled with antibodies against Clarias α,β-gonadotropin (GTH) and salmon G100-GTH. Only the large basophils were immunolabeled with anti-carp β-GTH, whereas the small basophils were the only cells immunolabeled with anti-human thyrotropin beta subunit (anti-h TSH-β). It was concluded that the large basophils represent the gonadotrops and the small basophils the thyrotrops. At the ultrastructural level the immunostaining of the GTH-cells was confined to three types of inclusions: (i) secretory vesicles, (ii) globules, and (iii) electron-dense, membrane-bound irregular masses. Especially the protein A- gold method, in combination with the use of a highly diluted homologous antiserum, resulted in a distinct localization of GTH. The presence of two types of nerve fibres, synaptically contacting the gonadotrops, is discussed with regard to the presence of a peptidergic (stimulatory) and an aminergic (inhibitory) control of GTH-secretion.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-0878
    Keywords: Liver ; Hepatocytes ; Ultrastructure ; Estradiol ; Vitellogenesis ; Teleosts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Hepatocytes of male zebrafish, Brachydanio rerio, were studied by means of light- and electron-microscopy, following a period of maximally 16 days of in-vivo treatment with estradiol-17β. The responsiveness of the male hepatocytes to this female sex steroid was investigated by use of morphometric methods. The results of this investigation show that the responsiveness was most obvious between 2 and 16 days, as revealed by an increase in cell size, accompanied by a proliferation of the granular endoplasmic reticulum and the Golgi complex. In addition, accumulations of glycogen granules, which are characteristic of hepatocytes in untreated males, had disappeared and lipid droplets had accumulated. These experimentally induced changes in the morphology of the male hepatocyte closely resemble those described for the female hepatocyte during the sexual cycle. It is concluded that the hepatocytes of male zebrafish can be stimulated by estradiol-17β to produce vitellogenin and that in female zebrafish this steroid is a key sex hormone responsible for vitellogenin production by the liver during the natural sexual cycle.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-0878
    Keywords: Gonadotropic cells ; LHRH analogue ; Pimo-zide ; Ultrastructure ; Goldfish ; Carassius auratus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effect of pimozide and an LHRH-analogue (LHRH-A) on gonadotropic cells of the goldfish pituitary gland were described qualitatively and quantitatively. A scale of four categories was devised to reflect various ultrastructural appearances of the cells. Experimental animals were divided into a control group, a group injected with LHRH-A alone, pimozide alone, and groups receiving these two substances in combination. Fish injected with the single substance were killed 12 h after injection while the groups receiving the combined treatments were killed at 4, 12 and 48 h. Serum levels of gonadotropin measured by radioimmunoassay were used to indicate whether an increase in hormone release had occurred. An immunocytochemical technique, the protein A-gold procedure, assured that the cells studied were gonadotropes. The control group showed variation in the profiles of gonadotropic cells. The single treatment groups showed some increase in secretory inclusions. At 4 h after injection the combined treatment caused a significant increase in hormone granules; at 12 and 48 h there was a gradual decrease in content of secretory products, and an increase in vacuolization. The results indicate that the combined pimozide and LHRH-A treatment stimulated gonadotropin production as well as release.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-0878
    Keywords: Pheromones ; Olfactory system ; Nervus terminalis ; Horseradish peroxidase ; Tracer studies ; Clarias gariepinus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The olfactory tract of the African catfish, Clarias gariepinus, consists of two tracts, the medial and lateral olfactory tract. Ovulated female catfish are attracted by male steroidal pheromones. Attraction tests with catfish in which the medial and lateral olfactory tract have been selectively lesioned show that the effects of these pheromones are mediated by the medial olfactory tract. The central connections of the medial and lateral olfactory tract have been studied by retro- and anterograde transport techniques using horseradish peroxidase as a tracer. Upon entering the forebrain, the medial olfactory tract innervates the posterior pars ventralis and pars supracommissuralis of the area ventralis telencephali and the nucleus preopticus periventricularis, the nucleus preopticus and the nucleus recessus posterioris. Application of horseradish peroxidase to the olfactory epithelium shows that part of the innervation of the area ventralis telencephali and the nucleus preopticus periventricularis can be attributed to the nervus terminalis, which appears to be embedded in the medial olfactory tract. The lateral olfactory tract sends projections to the same brain areas but also innervates the nucleus habenularis and a large terminal field in the area dorsalis telencephali pars lateralis ventralis. Furthermore, the medial olfactory tract carries numerous axons from groups of perikarya localized in the area dorsalis telencephali. Contralateral connections have been observed in the olfactory bulb, telencephalon, diencephalon and mesencephalon. It is suggested that processes of the medial olfactory tract innervating the preoptic region may influence the gonadotropin-releasing hormone system and in doing so may lead to behavioral and physiological changes related to spawning.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1573-5168
    Keywords: Brachydanio rerio ; Clarias gariepinus ; teleosts ; ovary ; testis ; seminal vesicles ; steroidogenesis ; steroid glucuronides ; sex pheromones
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In general, female zebrafish,Brachydanio rerio, ovulate only in the presence of males. The stimulant must be pheromonal as even male holding water is capable of inducing ovulation. After ovulation the mating phase begins. During this phase the male follows the female and oviposition as well as fertilization takes place. Both the ovulation and the mating are controlled by pheromones synthesized by the gonads. Ovulation can be induced by testicular homogenates. After the lipid material has been extracted from the testicular homogenates, the remaining aqueous phase can still induce ovulation. However, when the aqueous phase is treated with the enzymeβ-glucuronidase, it loses the ability to induce ovulation. This is an indication that glucuronides, probably steroid glucuronides, are the compounds responsible. During the mating phase, ovulated female zebrafish become attractive to males. It was found that, after ovulation, ovarian extracts contain the compounds responsible for attracting males. The attractant consists of a mixture of steroid glucuronides. After incubation of the gonads with3H-precursors seven steroid glucuronides have been identified in the testis and five in the ovary. Under fish culture conditions the African catfish,Clarias gariepinus, can produce postivitellogenic oocytes throughout the year. However, in capitivity neither males nor females spawn. In female catfish maturation and ovulation can be induced by treatment with gonadotropins. It might be possible that, analogous to the zebrafish, some reproductive processes in the catfish have to be induced by pheromones. It has been demonstrated that pheromonal compounds released by the seminal vesicles are involved in the attraction of female conspecifics. The steroid glucuronide synthesizing capability of the testes and the seminal vesicles of the male catfish are examined, as well as that of the ovary before and after ovulation of the female catfish. Both testes and seminal vesicles appear to be capable of steroid biosynthesis but only the latter synthesizes steroid glucuronides. Six of these conjugates have been isolated and identified. In the female catfish the ovaries are capable of synthesizing seven steroid glucuronides, but only after ovulation.
    Type of Medium: Electronic Resource
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  • 9
    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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  • 10
    ISSN: 1573-5168
    Keywords: oocyte maturation ; ovulation ; steroidogenesis ; teleosts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Changes in ovarian steroidogenesis accompanying oocyte maturation and ovulation were studied in the African catfish,Clarias gariepinus. Laboratory-reared females with postvitellogenic ovaries were treated with pimozide and LHRH-analogue. The plasma gonadotropin levels were determined by means of a homologous radioimmunoassay, the condition of the ovaries was studied by histological examination of the follicles, and the steroidogenetic capacity of the ovaries was analyzed byin vitro incubation of tissue fragments for 3 h with [3H]-pregnenolone and [3H]androstenedione as precursors. Data were collected at regular intervals between 0 and 16 h after pimozide-LHRH analogue administration. Until 4 h after the beginning of the experiments the plasma gonadotropin levels did not rise above 25 ng/ml, the ovaries remained in the stage of postvitellogenesis, and testosterone was the main end product of steroidogenesis. Four hours later the gonadotropin concentration in the blood had risen to more than 150 ng/ml, and the ovaries had entered the stage of germinal vesicle migration. At the same time steroidogenesis shifted towards the production of 17α,20β-dihydroxy-4-pregnen-3-one, 5β-pregnane-3α, 17α-diol-20-one, 5β-pregnane-3α,6α,17α-triol-20-one, 5β-pregnane-3α,17α,20β-triol and 5β-pregnane-3α,6α,17α,20β-tetrol. During the subsequent stage of germinal vesicle breakdown the plasma gonadotropin level remained high, and the synthesis of the C21-steroids showed a further increase. Simultaneously, the production of some C19-steroid glucuronides was enhanced. The preovulation and especially the postovulation stages were accompanied by a gradual decrease in steroidogenic capacity of the ovaries, even though the plasma gonadotropin level remained high. It is concluded that the prematuration surge of gonadotropin influences the activity of enzymes involved in steroidogenesis, leading to a reduced C17,20-lyase and to an augmented activity of the enzymes 20β-hydroxysteroid dehydrogenase (HSD), 5β-reductase, 3α-HSD, 6α-hydroxylase and UDP-glucuronosyltransferase. During ovulation the activity of all steroidogenic enzymes, including such key enzymes as 3β-HSD and 17α-hydroxylase, gradually decreases. Not only 17α,20β-dihydroxy-4-pregnen-3-one, but also the 5β-reduced pregnanes may be involved in inducing oocyte maturation and/or ovulation. The very polar triol and tetrol products may function, together with the steroid glucuronides as sex pheromones.
    Type of Medium: Electronic Resource
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