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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    Journal of neuroendocrinology 10 (1998), S. 0 
    ISSN: 1365-2826
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Dopamine D2-receptor activation directly inhibits GnRH-induced gonadotropin-II (maturational gonadotropin, GTH-II) secretion from goldfish pituitary cells. In this study, we show that dopamine and its D2 agonist, quinpirole, reduced GTH-II secretion induced by either high extracellular K+ concentration or the voltage-gated Ca2+ channel agonist, Bay K 8644. These actions of dopamine were blocked by addition of the dopamine D2-receptor antagonist, spiperone. The actions of dopamine on Ca2+ current in single identified goldfish gonadotrophs were assessed in voltage-clamp experiments using Ba2+ as the charge carrier through voltage-gated Ca2+ channels. Dopamine caused a concentration-dependent reduction in Ba2+ current amplitude with an EC50 of 1.0±0.3 nM, but did not shift the current-voltage relationship. The D2 agonist quinpirole also caused a dose-dependent reduction in the Ba2+ current amplitude with an EC50 of 2.7±1.4 nM. Quinpirole slowed the activation and inactivation kinetics, as well as removing the steady-state inactivation properties of the Ba2+ current. In contrast to the actions of quinpirole, the dopamine D1-receptor agonist, SKF 38393, did not affect the Ba2+ current. The inhibitory action of dopamine on voltage-dependent Ca2+ currents was reversed by spiperone, but not by the D1 antagonist SKF 83566. Voltage-dependent Na+ and K+ currents were not affected by dopamine or dopamine agonists. These data indicate that dopamine D2-receptor activation reduces Ca2+ influx through voltage-dependent Ca2+ channels to inhibit GTH-II secretion.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    Journal of neuroendocrinology 9 (1997), S. 0 
    ISSN: 1365-2826
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: In goldfish, gonadotropin-releasing hormone (GnRH) stimulation of growth hormone (GH) release has been shown to involve extracellular Ca2+ entry through voltage-sensitive Ca2+ channels and the activation of protein kinase C (PKC). In this study, the possible involvement of extracellular Na+ in mediating the GH response to GnRH was examined using dispersed pituitary cells. Perifusion with Na+-depleted medium reversibly reduced the acute GH response to 5-min pulses of either 10 nM salmon (s)GnRH or 10 nM chicken (c)GnRH-II. Similarly, replacement of normal medium with Na+-depleted medium attenuated the long-term GH release response to sGnRH and cGnRH-II under static incubation conditions. These results suggest that GnRH-induced GH release requires the presence of extracellular Na+. Treatment with 5-min pulses of the Na+-channel agonist veratridine (10 μM) increased GH release in an extracellular Ca2+-dependent manner, presumably due to activation of voltage-sensitive Ca2+ channels resulting from the depolarizing effect of increased Na+ influx. On the other hand, Na+ entry through tetrodotoxin (TTX)-sensitive, voltage-dependent Na+ channels is not involved in GnRH-induced GH release. Application of 250 nM TTX, which abolished the voltage-sensitive Na+ currents in identified goldfish somatotropes, did not affect the acute GH responses to 5-min pulses of sGnRH and cGnRH-II. The possible participation of Na+/H+ antiport in mediating the extracellular Na+-dependent GnRH action on GH release was then examined. In static incubation experiments, sGnRH- and cGnRH-II-induced GH secretion were reduced by inhibitors of the Na+/H+ antiport, amiloride and dimethylamiloride (DMA). Likewise, the GH response to the PKC activator, tetradecanoyl phorbol acetate, was attenuated by treatment with Na+-depleted medium, amiloride, and DMA. The inhibitory actions of amiloride and DMA were selective as these drugs did not affect the GH response elicited by the Ca2+ ionophore ionomycin and the voltage-sensitive Ca2+ channel agonist, Bay K 8644. Taken together, these results indicate that extracellular Na+ and the Na+/H+ exchanger are involved in the mediation of GnRH-stimulated GH release in goldfish. Furthermore, this dependence on Na+ and Na+/H+ antiport probably occurs distal to the activation of PKC by GnRH.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0878
    Keywords: Cell identification ; Cell separation ; Gonadotropin cells ; Prolactin cells ; Growth-hormone cells ; Immunofluorescence ; Differential interferencecontrast (Nomarski) microscopy ; Goldfish (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract To better understand neuroendocrine regulation and the intracellular mechanisms mediating pituitary-hormone release, it is necessary to study the physiology of identified single cells. We have developed a system to identify gonadotropin, growth-hormone, and prolactin cells in primary cultures of goldfish pituitary cells. Using Nomarski differential interference-contrast microscopy, the unique morphologies of discrete subpopulations of cells were characterized. To aid in the initial characterization of different pituitary-cell types, a discontinuous Percoll density-gradient cell-separation technique was developed. This method provided fractions enriched with functional gonadotropin, growth-hormone, and prolactin cells. The morphology of each cell type was initially characterized in enriched fractions of immunofluorescently labelled cells using differential interference-contrast microscopy. The cell type-specific morphologies were then confirmed in live pituitary-cell cultures. Gonadotropin, growth-hormone, and prolactin cells were correctly identified in live pituitary-cell cultures. Gonadotropin, growth-hormone, and prolactin cells were correctly identified in live mixed cultures in 92, 94, and 100% of the trials, respectively. The ability to directly identify cells in primary cultures allows the physiological study of identified single cells with minimal pretreatment.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-0878
    Keywords: Key words: Cell identification – Cell separation – Gonadotropin cells – Prolactin cells – Growth-hormone cells – Immunofluorescence – Differential interference-contrast (Nomarski) microscopy – Goldfish (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. To better understand neuroendocrine regulation and the intracellular mechanisms mediating pituitary-hormone release, it is necessary to study the physiology of identified single cells. We have developed a system to identify gonadotropin, growth-hormone, and prolactin cells in primary cultures of goldfish pituitary cells. Using Nomarski differential interference-contrast microscopy, the unique morphologies of discrete subpopulations of cells were characterized. To aid in the initial characterization of different pituitary-cell types, a discontinuous Percoll density-gradient cell-separation technique was developed. This method provided fractions enriched with functional gonadotropin, growth-hormone, and prolactin cells. The morphology of each cell type was initially characterized in enriched fractions of immunofluorescently labelled cells using differential interference-contrast microscopy. The cell type-specific morphologies were then confirmed in live pituitary-cell cultures. Gonadotropin, growth-hormone, and prolactin cells were correctly identified in live mixed cultures in 92, 94, and 100bad switch yylook 1 of the trials, respectively. The ability to directly identify cells in primary cultures allows the physiological study of identified single cells with minimal pretreatment.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Fish physiology and biochemistry 11 (1993), S. 77-84 
    ISSN: 1573-5168
    Keywords: goldfish ; pituitary ; growth hormone ; gonadotropin ; somatostatin ; dopamine ; D1 receptors ; body growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Description / Table of Contents: Résumé Des approches menées in vivo et in vitro ont été utilisées pour examiner le rôle de la dopamine (DA) en tant que facteur stimulant la libération d'hormone de croissance (GH) chez le poisson rouge. La DA stimule la libération de GH d'une façon dose dépendante par des fragments hypophysaires maintenus en périfusion. L'action stimulatrice de DA sur la libération de GH varie avec la saison avec un effet maximal chez les poissons sexuellement régressés, intermèdiare chez les poissons en recrudescence et minimal chez les animaux matures (préreproduction). La résponse en GH à la DA est bloquée par l'antagoniste des récepteurs dopaminergiques D1 (+)SCH23390, confirmant l'implication de ces types de récepteurs dans la libération de la GH induite par la DA. En utilisant des incubations statiques de cellules hypophysaires, la somatostatine, inhibiteur connu de la libération de GH chez le poisson rouge, abolit la réponse en GH à la DA. Des injections intrapéritonèales d'apomorphine, agoniste non sélectif de la dopamine, augmente les teneurs en GH plasmatique et la croissance linèaire du poisson rouge. Ces résultats suggèrent fortement le rôle de la DA comme facteur stimulant la libération de GH par l'intermèdiaire de récepteurs dopaminergiques D1 chez le poisson rouge.
    Notes: Abstract In vivo and in vitro approaches have been used to examine the role of dopamine (DA) as a growth hormone (GH)-releasing factor in the goldfish. DA stimulated GH release from perifused pituitary fragments of goldfish in a dose-dependent manner. The GH-releasing effect of DA was seasonal, being the highest in sexually regressed fish, intermediate in recrudescent fish, and the lowest in sexually mature (prespawning) fish. The GH response to DA was blocked by the D1 antagonist (+)SCH23390, confirming the involvement of D1 receptors in DA-stimulated GH release. In studies using static incubation of pituitary cells, somatostatin, a known physiological GH-release inhibitor in the goldfish, abolished the GH response to DA. Intraperitoneal injection of apomorphine, a non-selective DA agonist, also increased the plasma GH levels and enhanced the linear body growth of goldfish. These results strongly suggest that DA, by acting through DA D1 receptors, functions as a GH-releasing factor in the goldfish.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Fish physiology and biochemistry 11 (1993), S. 25-33 
    ISSN: 1573-5168
    Keywords: signal transduction ; goldfish gonadotropin ; goldfish growth hormone ; GnRH ; dopamine D1 and D2 activation ; calcium ; PKC ; cAMP ; Arachidonic acid ; G-proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Description / Table of Contents: Résumé Cette revue présente les données expérimentales démontrant l'implication de Ca++, de la protéine kinase C et du métabolismes de l'acide arachidonique dans les mécanismes régulant la sécrétion des hormones gonadotrope (GTH) et de croissance (GH). Des modèles de signaux de transduction de l'action de la gonadolibérine (GnRH) et de la dopamine sur la sécrétion de GTH et de GH sont proposés. Les deux GnRHs existant chez le poisson rouge pourraient se lier au même type de récepteur et activer différentes voies de transduction dans deux différents types cellulaires (GTH vs. GH) ou dans un seul type (GTH).
    Notes: Abstract Evidence for the involvement of Ca2+, protein kinase C, cAMP, and arachidonic acid metabolism in mediating gonadotropin (GTH) and growth hormone (GH) release in the goldfish is reviewed. Models for the signal transduction pathways mediating GTH-releasing hormone (GnRH) and dopamine actions on GTH and GH secretion are postulated. A novel hypothesis that two GnRHs which bind to the same receptor type activate different transduction cascade in two different cell types (GTH vs. GH) as well as within the same cell type (GTH) is presented.
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  • 7
    ISSN: 1573-5168
    Keywords: PKC down regulation ; GnRH action ; dopamine D2 action ; goldfish pituitary cells ; static culture ; perifusion experiments
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Description / Table of Contents: Résumé Il a été établi que chez le poisson rouge, les sécrétions de gonadotropine (GtH) et d'hormone de croissance (GH) sont toutes les deux stimulées par la gonadolibérine (GnRH); de plus, la sécrétion de GtH est inhibée par des mécanismes dopaminergiques de type D2. Dans le présent travail, la déplétion de la teneur en protéine kinase C (PKC) dans des cellules hypophysaires de poisson rouge réduit les résponses en GtH et GH au GnRH et à un activateur de la PKC de cellules maintenues en incubation statique. Dans des cellules maintenues en périfusion et soumises à une déplétion en PKC, la GtH libérée en réponse à un analogue du sGnRH est fortement diminuée, cependent les réponses hormonales à la forskoline sont augmentées. La stimulation des récepteurs dopaminergiques D2 réduit, dans le cas d'action d'activateur de la PKC, la réponse en GtH mais pas en GH. Ces résultats indiquent que la PKC est impliquée dans les mécanismes de régulation de GtH et GH par des facteurs neuroendocriniens naturels.
    Notes: Abstract It has been established that secretion of gonadotropin (GtH) and growth hormone (GH) release in goldfish are both stimulated by GtH-releasing hormone (GnRH); in addition GtH secretion is inhibited by dopamine D2 mechanisms. In the present study, depletion of protein kinase C (PKC) in goldfish pituitary cells reduced the GtH and GH responses to GnRH and an activator of PKC in static culture. In perifusion studies, GtH released in response to sGnRH analog was greatly attenuated in PKC-depleted cells, however, hormone responses to forskolin were enhanced. Stimulation of dopamine D2 receptors reduced the GtH, but not the GH, responses elicited by PKC activators. These results indicate that PKC participates in the GtH and GH responses to natural neuroendocrine regulators in the goldfish.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1573-5168
    Keywords: signal transduction ; cyclic AMP ; protein kinase A ; protein kinase C ; [Ca2+]i ; voltage-sensitive Ca2+ channels
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The mechanisms of pituitary adenylate cyclase activating polypeptide (PACAP) action on goldfish growth hormone (GH) release were investigated by examining GH release responses from dispersed goldfish pituitary cells to a synthetic mammalian (m)PACAP38 peptide. It was established that GH release stimulated by 2-h exposure to mPACAP38 was concentration-dependent, attenuated by the PACAP receptor antagonist mPACAP6−38, and subject to neuroendocrine modulation by somatostatin. Maximal mPACAP38-stimulated GH release was not additive to the responses elicited by either the adenylate cyclase activator forskolin or the cyclic (c)AMP analog 8-bromo-cAMP. The GH responses to mPACAP38, forskolin and 8-bromo-cAMP, either alone or in combination, were abolished by H89, a protein kinase A (PKA) inhibitor. SQ22536, an adenylate cyclase inhibitor, attenuated forskolin- and mPACAP38-stimulated GH release. In contrast, mPACAP38-stimulated GH release were additive to the responses to two protein kinase C (PKC) activators and unaffected by two PKC inhibitors. These results suggest that the stimulatory action of PACAP on GH secretion is mediated through a cAMP- / PKA-dependent mechanism, whereas the involvement of PKC appears unlikely. The ability of mPACAP38 to further enhance maximal GnRH (PKC)-dependent GH release, but not dopamine D1 agonist (PKA)-dependent GH secretion, is consistent with this hypothesis. A possible involvement of Ca2+ in PACAP action is also suggested. Two inhibitors of voltage-sensitive Ca2+ channel reduced the GH responses to mPACAP38 in static incubation; conversely, mPACAP38 increased intracellular [Ca2+] in identified, single goldfish somatotropes.
    Type of Medium: Electronic Resource
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