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Interaction of cyclic AMP- and calcium-dependent mechanisms in the regulation of growth hormone-releasing hormone-stimulated growth hormone release from ovine pituitary cells



Interaction of cyclic AMP- and calcium-dependent mechanisms in the regulation of growth hormone-releasing hormone-stimulated growth hormone release from ovine pituitary cells



Domestic Animal Endocrinology 13(3): 229-238



Growth hormone-releasing hormone (GHRH)-stimulated growth hormone (GH) release from the sheep pituitary is mediated through Ca(2+)-and cyclic AMP-dependent mechanisms. The initial Ca2+ influx is suggested to result from depolarization, whereas a secondary Ca2+ influx is thought to result from second messengers. This study sought to determine whether there was an interaction between these two signal transduction pathways. Sheep pituitary cells were placed in culture for 4 d and were then washed and incubated for 1 hr in serum-free medium before the application of specific antagonises and/or agonists. Both KCl and forskolin stimulated GH release (P < 0.05), but neither produced an effect similar to that of GHRH. The combination of both stimuli, however, mimicked GH release, as seen with a maximal dose of GHRH. Pretreatment with H89 (protein kinase A [PKA] inhibitor) inhibited GHRH, forskolin- and KCl-stimulated GH release (P < 0.001) but had no effect on phorbol myristate acetate (PMA)-stimulated GH release. Verapamil (voltage-dependent Ca2+ channel blocker) inhibited the GHRH effects on GH release (P < 0.0002) but did not influence forskolin or PMA actions. These data suggest that Ca(2+)-dependent pathways converge with cyclic AMP-dependent pathways before or with the activation of PKA. The data also suggest that PKA activation by cyclic AMP alone is insufficient to reproduce either the effects of GHRH stimulation or the combined effects of Ca2+ influx plus PKA activation on GH release. A calmodulin blocker, W7, reduced GHRH-stimulated GH release, a reduction equivalent to the Ca2+ effect on GH release. This suggests that Ca2+ activates calmodulin, which in turn enhances adenylyl cyclase and/or PKA activity to release GH from the sheep pituitary.

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Accession: 046429863

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PMID: 8738864

DOI: 10.1016/0739-7240(95)00069-0



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