title: Growth hormone can regulate functions of porcine ovarian granulosa cells through the cAMP/protein kinase A system.
abstract: The action of growth hormone (GH) on the production of hormones, growth factors, growth factor binding protein and the occurrence of apoptosis in porcine ovarian granulosa cells, as well as the role of cAMP-stimulated protein kinase A (PKA) in the mediation of these effects, were studied. For this purpose, the effects of exogenous pGH (1-10,000 ng/ml), PKA blockers KT5720 (100 ng/ml) and Rp-cAMPS (1micromol), alone and in combination, on insulin-like growth factor type I (IGF-I), insulin-like binding protein 3 (IGFBP-3), oxytocin (OT) and prostaglandin F alpha (PGF) secretion, PKA and cAMP response element binding transcription factor (CREB) content and the occurrence of apoptosis were investigated. It was found (using RIA/IRMA) that GH addition to culture medium significantly stimulated IGF-I and PGF release and inhibited IGFBP-3 and OT secretion. GH significantly decreased the incidence of apoptosis (TUNEL method) in cultured cells. Immunocytochemical study and Western immunoblotting showed, that addition of GH caused a dramatic increase in the accumulation of immunoreactive PKA within the cells, whilst Western blotting did not reveal marked influence of GH on content of CREB in cell lysates. PKA blockers, given alone, were able to decrease IGFBP-3 output (Rp-cAMPS, but not KT5720), reduce basal OT release (either Rp-cAMPS and KT5720) and increase PGF accumulation (KT5720, but not Rp-cAMPS). Furthermore, PKA blockers were able to prevent stimulatory effects of GH on IGF-I and PGF release, and inhibitory effect of GH on IGFBP-3, OT output and on apoptosis. These observations suggest the involvement of GH and a PKA-dependent intracellular mechanism in the control of IGF-I, IGFBP-3, OT, PGF, cAMP and apoptosis in porcine ovarian granulosa cells. Stimulation of PKA by GH and the prevention of GH-induced effects by PKA blockers suggest that both stimulatory and inhibitory effects of GH on porcine ovarian cells are probably mediated by the cAMP/PKA system.
authors.affiliation: Research Institute of Animal Production, Nitra, Slovak Republic. sirotkin@vuzv.sk
authors.forename: A V
authors.initials: AV
authors.lastname: Sirotkin
authors.forename: A V
authors.initials: AV
authors.lastname: Makarevich
mesh_terms: D000818:Animals; D017209:Apoptosis; D002227:Carbazoles; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000502:physiology*; D017362:Cyclic AMP Response Element-Binding Protein / Q000032:analysis; D017868:Cyclic AMP-Dependent Protein Kinases / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006107:Granulosa Cells / Q000737:chemistry / Q000502:physiology*; D013006:Growth Hormone / Q000494:pharmacology*; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D007211:Indoles / Q000494:pharmacology; D018972:Insulin-Like Growth Factor Binding Protein 3 / Q000378:metabolism; D007334:Insulin-Like Growth Factor I / Q000378:metabolism; D010121:Oxytocin / Q000378:metabolism; D011460:Prostaglandins F / Q000378:metabolism; D011758:Pyrroles / Q000494:pharmacology; D013552:Swine / Q000502:physiology*
chemical_list: D002227:Carbazoles; D017362:Cyclic AMP Response Element-Binding Protein; D004791:Enzyme Inhibitors; D007211:Indoles; D018972:Insulin-Like Growth Factor Binding Protein 3; D011460:Prostaglandins F; D011758:Pyrroles; D010121:Oxytocin; C057416:KT 5720; D007334:Insulin-Like Growth Factor I; D013006:Growth Hormone; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases