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The aim of the study was to analyze the contribution of kisspeptin-10 (KiSS-10) and peptide 234 (kisspeptin-234, potent neutral antagonist of GPR-54 receptors) to the modulation of GnRH-induced folliclestimulating hormone (FSH) secretion from anterior pituitary cells of prepubertal ram lambs in vitro. Pituitary cells were cultured in McCoy 5A medium without hormones (the negative control), with GnRH (4 × 10⁻⁹ M, the positive control), with GnRH (4 × 10⁻⁹ M) and 10⁻¹¹-10⁻⁸ M of KiSS-10 or GnRH (4 × 10⁻⁹ M), 10⁻¹¹-10⁻⁸ M of KiSS-10 and 10⁻⁷ M of peptide 234. After 6, 12 and 48 h of the experiment, the secretion of follicle-stimulating hormone was determined. The obtained results show that FSH secretion from anterior pituitary cells of ram lambs in vitro was dependent on kisspeptin-10 concentration in the culture medium. Addition of 10⁻¹¹-10⁻⁹ M of KiSS-10 caused an increase in FSH secretion (r = 0.73, 0.90, and 0.82 after 6, 12 and 48 h, respectively) compared to both the negative and positive control, whereas the highest concentration of KiSS-10 (10⁻⁸ M) suppressed the secretion of this gonadotropin. The most stimulating effect was observed under the influence of 10⁻⁹ M of KiSS-10. However, concurrent cell exposure to peptide 234 abolished the stimulating action of kisspeptin-10 on FSH secretion. The negative correlation between FSH secretion and 10⁻¹¹-10⁻⁸ M of KiSS-10 in this condition was found (r = –0.68, –0.91, and –0.81 after 6, 12 and 48 h, respectively). This confirms that the observed increase in GnRH-induced FSH secretion was a direct effect of KiSS-10 on the anterior pituitary cells of prepubertal ram lambs.
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Cortistatin and pituitary hormone secretion in rat

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Cortistatin (CST), a novel neuropeptide, shows high structural homology and functional resemblance with somatostatin. CST binds with high affinity to all somatostatin receptors, and contrary to somatostatin, is also able to bind with MrgX2 and GH secretagogue receptor of ghrelin (GHS-R1) receptors. The aim of the present investigation was to evaluate in vivo the effect of peripheral administration of cortistatin on pituitary hormone release in comparison with somatostatin (SS) treatment. Adult male rats used in the experiment, were given peripheral injection of cortistatin, somatostatin or vehicle. Blood was withdrawn 60 and 120 minutes thereafter. We found short lasting significant decrease of GH concentration as a result of administration of CST and SS when compared with saline injected controls. Prolactin levels were increased 60 min after cortistatin but not to somatostatin injection. There was no effect of CST on both LH and FSH concentration; however, SS administration influenced gonadotropin secretion. We conclude that cortistatin play a regulatory role in pituitary secretion. Moreover, some differences have been found when compared cortistatin to somatostatin. Thus, when analyzing the mechanism of cortistatin activity it is worth to consider the effect of binding with receptors of somatostatin, specific receptor for CST (MrgX2) and GHS-R.
The objective of this study was to examine the degradation of short peptides corre­sponding to modified fragments of human growth hormone-releasing hormone by trypsin. Six analogues of pentapeptide 9-13 of human growth hormone-releasing hor­mone containing homoarginine, ornithine, glutamic acid, glycine, leucine or phenyl­alanine residue in position 11, two analogues of hexapeptide 8-13 of human growth hormone-releasing hormone and two analogues of heptapeptide 7-13 of human growth hormone-releasing hormone containing homoarginine or glycine residue in position 11 were obtained. The peptides were subjected to digestion by trypsin and the course of reaction was monitored using HPLC. It was found that the rate of hy­drolysis of the Lys12-Val13 peptide bond depends on the amino-acid residue preceding Lys12 . The extension of the peptide chain towards the N-terminus by introduction of consecutive amino-acid residues corresponding to the human growth hormone-re­leasing hormone sequence accelerates the hydrolysis process. These results may be of assistance in designing new analogues of human growth hormone-releasing hor­mone, more resistant to the activity of proteolytic enzymes.
Background. Ghrelin, a newly discovered hormone is involved mainly in the regulation of body energy homeostasis. It has also been proved that ghrelin affects many other processes including the control of the hypothalamo-pituitary-gonadal axis in vertebrates. Ghrelin interacts also with other peptides and neurotransmitters, which are involved in gonadotropin release, such as endogenous opioid peptides. The aim of the experiment was to compare the effects of ghrelin and naltrexone, an opioid antagonist, on LH secretion by pituitary cells of mature female and male common carp, Cyprinus carpio L. Materials and methods. In the in vitro experiment the time-dependent (10 and 24 h) action of ghrelin (10-7 or 10-6 M) and naltrexone at 10-6 M added alone or in combination on LH secretion by enzymatically dispersed pituitary cells of mature female and male carp (Cyprinus carpio L.) was studied. Results. Ghrelin alone at a dose of 10-6 M stimulated LH secretion after 10 h of female- but not male-derived cell incubations. Ghrelin at 10-7 M had no effect on LH secretion from either female or male cells. Naltrexone alone stimulated LH secretion only after 24 h of female cell culture. In male cells no significant changes in LH secretion in response to naltrexone alone were found after 10 or 24 h incubation period. Combined treatment (ghrelin 10-7 or 10-6 M and naltrexone) stimulated LH secretion in female and male cell incubations at 10 and 24 h: LH levels were significantly higher in comparison to control, to ghrelin alone (in cells of both sexes) and to naltrexone alone (in male cells only). Conclusion. The results suggest that opioids and ghrelin may control LH secretion in carp acting synergistically, probably through the same receptor type. The concept of opioid and ghrelin interaction in the gonadal steroid feedback on LH release is also discussed.
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