Growth hormone can regulate functions of porcine ovarian granulosa cells through the cAMP/protein kinase A system

We have studied the action of GH on the production of hormones, growth factors, growth factor-binding protein and the occurrence of apoptosis in bovine ovarian granulosa cells, as well as the role of cAMP-stimulated protein kinase A (PKA) in the mediation of these effects. For this purpose we investigated the effects of exogenous bovine GH (0.001-10 microgram/ml), PKA blockers KT5720 (100 ng/ml) and adenosine-3',5'-monophosphothiodate (Rp-cAMPS) (1 micromol), alone and in combination, on IGF-I, IGF-binding protein (IGFBP)-3, oxytocin, progesterone and estradiol secretion, cAMP and PKA content and the occurrence of apoptosis.The secretion of hormones, IGF-I and IGFBP-3 into the culture medium was measured using RIA/IRMA. The presence of PKA was detected using immunocytochemistry and Western immunoblotting. The presence of cAMP in cells was demonstrated using immunocytochemistry, whilst the proportion of apoptotic cells was determined by the TUNEL method.It was found that the addition of GH to the culture medium strongly (P<0.05) stimulated IGF-I (at a concentration of 0.001-10 microgram GH/ml medium), IGFBP-3 (0.001-1 microgram GH/ml) and oxytocin (0.01-10 microgram GH/ml) secretion. Low concentrations (1-100 ng/ml) of GH stimulated, whilst a higher concentration (10 microgram/ml) inhibited estradiol output. GH slightly (P<0.05) inhibited progesterone (1-100 ng GH/ml) secretion and significantly (P<0.05) decreased the incidence of apoptosis (0.01-1 microgram GH/ml) in cultured cells. The addition of GH (100 ng/ml) caused a dramatic (P<0.05) increase in the proportion of cells possessing the immunoreactive catalytic subunit of PKA and a slight decrease in the proportion of cells containing the regulatory PKA subunit.PKA blockers KT5720 and Rp-cAMPS significantly (P<0.05) reduced the proportion of granulosa cells containing cAMP, and the catalytic and (in the case of KT5720) regulatory subunits of PKA. KT5720 given alone significantly (P<0.05) inhibited the secretion of IGFBP-3, but not that of IGF-I or progesterone. Rp-cAMPS decreased (P<0.05) the secretion of oxytocin but not that of estradiol output or the occurrence of apoptosis. KT5720 and Rp-cAMPS fully or partially prevented the GH effect on IGF-I, IGFBP-3, oxytocin, progesterone, estradiol and apoptosis.These observations suggest the involvement of GH and a cAMP/PKA-dependent intracellular cascade in the control of IGF-I, IGFBP-3, oxytocin, progesterone, estradiol, cAMP and apoptosis in bovine ovarian granulosa cells. The stimulation of PKA by GH and the prevention of GH-induced effects by PKA blockers suggest that the observed GH effects on bovine ovarian cells are probably mediated by the cAMP/PKA system.

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The role of GH/IGF-I/cAMP/protein kinase a axis in the control of apoptosis and secretory activity of bovine ovarian granulosa cells

Growth Hormone & IGF Research ◽

10.1016/s1096-6374(98)80216-5 ◽

1998 ◽

Vol 8 (4) ◽

pp. 333

Author(s):

A Sirotkin ◽

A Makarevich ◽

P-G Marnet ◽

L Hetenyi ◽

J Bulla

Keyword(s):

Protein Kinase ◽

Protein Kinase A ◽

Granulosa Cells ◽

Secretory Activity ◽

Ovarian Granulosa Cells ◽

Igf I ◽

Kinase A

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Follicle-stimulating Hormone Stimulates Protein Kinase A-mediated Histone H3 Phosphorylation and Acetylation Leading to Select Gene Activation in Ovarian Granulosa Cells

Journal of Biological Chemistry ◽

10.1074/jbc.m106710200 ◽

2001 ◽

Vol 276 (43) ◽

pp. 40146-40155 ◽

Cited By ~ 114

Author(s):

Lisa M. Salvador ◽

Youngkyu Park ◽

Joshua Cottom ◽

Evelyn T. Maizels ◽

Jonathan C. R. Jones ◽

...

Keyword(s):

Protein Kinase ◽

Protein Kinase A ◽

Granulosa Cells ◽

Follicle Stimulating Hormone ◽

Gene Activation ◽

Histone H3 ◽

Ovarian Granulosa Cells ◽

H3 Phosphorylation ◽

Histone H3 Phosphorylation ◽

Kinase A

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Luteinizing Hormone Receptor Activation in Ovarian Granulosa Cells Promotes Protein Kinase A-Dependent Dephosphorylation of Microtubule-Associated Protein 2D

Molecular Endocrinology ◽

10.1210/me.2007-0457 ◽

2008 ◽

Vol 22 (7) ◽

pp. 1695-1710 ◽

Cited By ~ 22

Author(s):

Maxfield P. Flynn ◽

Evelyn T. Maizels ◽

Amelia B. Karlsson ◽

Thomas McAvoy ◽

Jung-Hyuck Ahn ◽

...

Keyword(s):

Protein Kinase ◽

Luteinizing Hormone ◽

Protein Kinase A ◽

Granulosa Cells ◽

Hormone Receptor ◽

Receptor Activation ◽

Luteinizing Hormone Receptor ◽

Ovarian Granulosa Cells ◽

Kinase A

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A-Kinase anchoring protein-13 (AKAP13) may be required for protein kinase A (PKA)-mediated aromatase Expression in granulosa cells

Fertility and Sterility ◽

10.1016/j.fertnstert.2017.07.328 ◽

2017 ◽

Vol 108 (3) ◽

pp. e107

Author(s):

K. C. Cayton Vaught ◽

P. Driggers ◽

J. Segars

Keyword(s):

Protein Kinase ◽

Protein Kinase A ◽

Granulosa Cells ◽

Aromatase Expression ◽

Anchoring Protein ◽

Kinase A

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Pituitary adenylate cyclase activating polypeptide as a novel hypophysiotropic factor in fish

Biochemistry and Cell Biology ◽

10.1139/o00-055 ◽

2000 ◽

Vol 78 (3) ◽

pp. 329-343 ◽

Cited By ~ 63

Author(s):

Anderson OL Wong ◽

Wen Sheng Li ◽

Eric KY Lee ◽

Mei Yee Leung ◽

Lai Yin Tse ◽

...

Keyword(s):

Growth Hormone ◽

Protein Kinase C ◽

Protein Kinase ◽

Adenylate Cyclase ◽

Protein Kinase A ◽

Anterior Pituitary ◽

Kinase C ◽

Pituitary Adenylate Cyclase ◽

Pacap Receptors ◽

Kinase A

Pituitary adenylate cyclase activating polypeptide (PACAP) is a novel member of the secretin-glucagon peptide family. In mammals, this peptide has been located in a wide range of tissues and is involved in a variety of biological functions. In lower vertebrates, especially fish, increasing evidence suggests that PACAP may function as a hypophysiotropic factor regulating pituitary hormone secretion. PACAP has been identified in the brain-pituitary axis of representative fish species. The molecular structure of fish PACAP is highly homologous to mammalian PACAP. The prepro-PACAP in fish, however, is distinct from that of mammals as it also contains the sequence of fish GHRH. In teleosts, the anterior pituitary is under direct innervation of the hypothalamus and PACAP nerve fibers have been identified in the pars distalis. Using the goldfish as a fish model, mRNA transcripts of PACAP receptors, namely the PAC1 and VPAC1 receptors, have been identified in the pituitary as well as in various brain areas. Consistent with the pituitary expression of PACAP receptors, PACAP analogs are effective in stimulating growth hormone (GH) and gonadotropin (GTH)-II secretion in the goldfish both in vivo and in vitro. The GH-releasing action of PACAP is mediated via pituitary PAC1 receptors coupled to the adenylate cyclase-cAMP-protein kinase A and phospholipase C-IP3-protein kinase C pathways. Subsequent stimulation of Ca2+ entry through voltage-sensitive Ca2+ channels followed by activation of Ca2+-calmodulin protein kinase II is likely the downstream mechanism mediating PACAP-stimulated GH release in goldfish. Although the PACAP receptor subtype(s) and the associated post-receptor signaling events responsible for PACAP-stimulated GTH-II release have not been characterized in goldfish, these findings support the hypothesis that PACAP is produced in the hypothalamus and delivered to the anterior pituitary to regulate GH and GTH-II release in fish.Key words: PACAP, VIP, PAC1 receptor, VPAC1 receptor, VPAC2 receptor, growth hormone, gonadotropin-II, cAMP, protein kinase A, protein kinase C, calcium, pituitary cells, goldfish, and teleost.

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Growth Hormone Releasing Peptide-2 Attenuation of Protein Kinase C-Induced Inflammation in Human Ovarian Granulosa Cells

International Journal of Molecular Sciences ◽

10.3390/ijms17081359 ◽

2016 ◽

Vol 17 (8) ◽

pp. 1359

Author(s):

Yi-Ning Chao ◽

David Sun ◽

Yen-Chun Peng ◽

Yuh-Lin Wu

Keyword(s):

Growth Hormone ◽

Protein Kinase C ◽

Protein Kinase ◽

Granulosa Cells ◽

Ovarian Granulosa Cells ◽

Kinase C

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Effect of Follicle-Stimulating Hormone and Estrogen on the Expression of Betaglycan Messenger Ribonucleic Acid Levels in Cultured Rat Granulosa Cells

Endocrinology ◽

10.1210/en.2004-1665 ◽

2005 ◽

Vol 146 (8) ◽

pp. 3379-3386 ◽

Cited By ~ 16

Author(s):

Yuki Omori ◽

Kazuto Nakamura ◽

Soichi Yamashita ◽

Hiroko Matsuda ◽

Tetsuya Mizutani ◽

...

Keyword(s):

Estrogen Receptor ◽

Protein Kinase ◽

Protein Kinase A ◽

Granulosa Cells ◽

Luciferase Assay ◽

Regulatory Function ◽

Type Ii ◽

Mrna Transcript ◽

Inhibin A ◽

Kinase A

Abstract Betaglycan (TGFβ type III receptor) was recently identified as a coreceptor to enhance the binding of inhibin A to activin type II receptor. This inhibin/betaglycan/activin type II receptor complex prevents activins from binding to their own receptors. The present study was undertaken to identify the expression and the regulation of the betaglycan gene in cultured rat granulosa cells. Northern blot analysis indicated betaglycan mRNA transcript of approximately 6.4 kbp. The treatment of the cells with FSH increased the betaglycan mRNA level, and a concurrent treatment with estradiol brought a significant increase in betaglycan mRNA. The protein kinase A activator, 8-bromoadenosine-cAMP, also increased the expression of its mRNA. Furthermore, betaglycan mRNA was induced additively by estradiol, which was blocked by estrogen receptor antagonists [ICI 182780, (R, R)-cis-diethyltetrahydro-2,8-chrysenediol]. In the luciferase assay, FSH altered the promoter activity of betaglycan. Moreover, when FSH plus estradiol was added to the granulosa cells, a significant increase in the half-life of betaglycan mRNA transcript was seen. In summary, FSH and estradiol increased betaglycan mRNA expression, most possibly through the protein kinase A pathway and the estrogen receptor-β. The increase of betaglycan mRNA was due to an increase in transcription and altered mRNA stability. In ovarian regulatory function, the expression of betaglycan may involve the functional antagonism of inhibin A in activin signal transduction.

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