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Exp Clin Endocrinol Diabetes 2004; 112(2): 108-114
DOI: 10.1055/s-2004-815766
Article

J. A. Barth Verlag in Georg Thieme Verlag Stuttgart · New York

The Role of Oxytocin, Protein Kinase A, and ERK-Related MAP-Kinase in the Control of Porcine Ovarian Follicle Functions

A. V. Makarevich1 , A. V. Sirotkin1 , J. Franek2 , H. B. Kwon3 , J. Bulla2
  • 1Research Institute of Animal Production, Nitra, Slovak Republic
  • 2State University of Agriculture, Nitra, Slovak Republic
  • 3Chonnam National University, Kwangju, Korea
Further Information

Publication History

Received: 20. März 2003 First decision: 28. April 2003

Accepted: 25. August 2003

Publication Date:
19 March 2004 (online)

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Abstract

The aim of our in vitro experiments was to study the role of oxytocin (OT), cAMP/protein kinase A (PKA), and mitogen-activated protein kinase (ERKs MAP-kinase) in the control of ovarian cell functions as well as the role of PKA and MAPK in mediating OT effects on these processes. The whole porcine ovarian follicles were cultured in the presence or absence of OT (1, 10, 100 ng/ml), PKA inhibitor Rp-cAMPS (10 nM), MAP-kinase inhibitor PD98059 (1 μg/ml), or their combination. The release of prostaglandins F (PGF) and E (PGE) were determined by RIA, PKA (α-cat subunit), the proliferation-associated peptide PCNA and ERK-1, -2 expression in cell lyzates were analysed by Western-blotting.

OT stimulated the release of PGF and PGE, and accumulation of PKA, ERK-1/-2, and PCNA in cell lysate. PD98059 decreased the basal PGF and PGE output, as well as reduced both ERK-1 and ERK-2 accumulation in cell lysates. Rp-cAMPS decreased PKA accumulation in cell lysates. Rp-cAMPS prevented the OT-induced stimulation of PKA, ERK-1, ERK-2, PGF, and PGE, PD98059 did so for PKA, PGF, and PGE. However, PD98059 reduced either basal or OT-induced p-ERK level. OT-stimulated PCNA accumulation was only slightly modified by these blockers.

These observations suggest that OT, PKA, and ERKs MAPK can be involved in the control of PGs release and proliferation of ovarian cells. The influence of OT on both PKA and MAPK, and the ability of PKA and MAPK blockers to prevent completely or partially OT effects suggest, that effects of OT on PGF and PGE can be mediated by both PKA and MAPK. The role of MAPK and PKA in mediating the proliferative effects of OT seems to be minor assuming the involvement of other intracellular messengers.

Key words

Porcine - ovarian follicle - oxytocin - prostaglandin - MAPK/ERKs - protein kinase A

References

  • 1 Ainsworth L, Tsang B K, Downey B R, Marcus G J. The synthesis and action of steroids and prostaglandins during follicular maturation in the pig.  J Reprod Fert. 1990;  40 137-150
    PubMedGoogle Scholar
  • 2 Burns P D, Mendes Jr J O. Yemm RS, Clay CM, Nelson SE, Hayes SH, Silvia WJ. Cellular mechanisms by which oxytocin mediates ovine endometrial prostaglandin F (2 alpha) synthesis: role of g (i) proteins and mitogen-activated protein kinases.  Biol Reprod. 2001;  65 1150-1155
    PubMedGoogle Scholar
  • 3 Cameron M R, Foster J S, Bukovsky A, Wimalasena J. Activation of mitogen-activated protein kinases by gonadotropins and cyclic adenosine 5′-monophosphates in porcine granulosa cells.  Biol Reprod. 1996;  55 111-119
    PubMedGoogle Scholar
  • 4 Cassoni P, Sapino A, Fortunati N, Munaron L, Chini B, Bussolati G. Oxytocin inhibits the proliferation of MDA-MB231 human breast-cancer cells via cyclic adenosine monophosphate and protein kinase A.  Int J Cancer. 1997;  72 340-344
    CrossrefPubMedGoogle Scholar
  • 5 Cassoni P, Sapino A, Munaron L, Deaglio S, Chini B, Graziani A, Ahmed A, Bussolati G. Activation of functional oxytocin receptors stimulates cell proliferation in human trophoblast and choriocarcinoma cell lines.  Endocrinol. 2001;  142 1130-1136
    CrossrefPubMedGoogle Scholar
  • 6 Chang K J, Kim J W, Lee J, Im W B, Kwon H B, Schuetz A W. Prostaglandin production and ovulation during exposure of amphibian ovarian follicles to gonadotropin or phorbol ester in vitro.  Gen Comp Endocrinol. 1995;  100 257-266
    CrossrefPubMedGoogle Scholar
  • 7 De Manno D A, Hunzicker-Dunn M. cAMP-dependent protein kinase isozymes in porcine follicles and corpora lutea.  Mol Cell Endocrinol. 1991;  80 91-104
    CrossrefPubMedGoogle Scholar
  • 8 Einspanier A, Jurdzinski A, Hodges J K. A local oxytocin system is a part of the luteinization process in the preovulatory follicle of the marmoset monkey (Callithrix jacchus).  Biol Reprod. 1997;  57 16-26
    PubMedGoogle Scholar
  • 9 Furuya K, Mizumoto Y, Makimura N, Mitsui C, Murakami M, Tokuoka S, Ishikawa N, Nagata I, Kimura T, Ivell R. A novel biological aspect of ovarian oxytocin: gene expression of oxytocin receptor in cumulus/luteal cells and the effect of oxytocin on embryogenesis in fertilized oocytes.  Adv Exp Med Biol. 1995;  395 523-528
    PubMedGoogle Scholar
  • 10 Keel B A, Davis J S. Epidermal growth factor activates extracellular signal-regulated protein kinases (ERK) in freshly isolated porcine granulosa cells.  Steroids. 1999;  64 654-658
    CrossrefPubMedGoogle Scholar
  • 11 Leung P, Steele G. Intracellular signalling in the gonads.  Endocr Rev. 1992;  13 476-498
    CrossrefPubMedGoogle Scholar
  • 12 Makarevich A V, Sirotkin A V. Presumptive mediators of growth hormone action on insulin-like growth factor I release by porcine ovarian granulosa cells.  Biol Sign Recept. 2000;  9 248-254
    PubMedGoogle Scholar
  • 13 Makarevich A V, Sirotkin A V, Chrenek P, Bulla J, Hetenyi L. The role of IGF-I, cAMP/protein kinase A and MAP-kinase in the control of steroid secretion, cyclic nucleotide production, granulosa cell proliferation and preimplantation embryo development in rabbits.  J Ster Biochem Mol Biol. 2000;  73 123-133
    PubMedGoogle Scholar
  • 14 Mayerhofer A, Sterzik K, Link H, Wiemann M, Gratzl M. Effect of oxytocin on free intracellular Ca2+ levels and progesterone release by human granulose lutein cells.  J Clin Endocrinol Metab. 1993;  77 1209-1214
    CrossrefPubMedGoogle Scholar
  • 15 Okuda K, Miyamoto A, Sauerwein H, Schweigert F J, Schams D. Evidence for oxytocin receptors in cultured bovine luteal cells.  Biol Reprod. 1992;  46 1001-1006
    PubMedGoogle Scholar
  • 16 Okuda K, Uenoyama Y, Fujita Y, Iga K, Sakamoto K, Kimura T. Functional oxytocin receptors in bovine granulosa cells.  Biol Reprod. 1997;  56 625-631
    PubMedGoogle Scholar
  • 17 Olsen M K, Reszka A A, Abraham I. KT5720 and U-98017 inhibit MAPK and alter the cytoskeleton and cell morphology.  J Cell Physiol. 1998;  176 525-536
    CrossrefPubMedGoogle Scholar
  • 18 Peng X, Maruo T, Matsuo H, Takekida S, Deguchi J. Serum deprivation-induced apoptosis in cultured porcine granulosa cells is characterized by increased expression of p53 protein, Fas antigen and Fas ligand and by decreased expression of PCNA.  Endocrine J. 1998;  45 247-253
    PubMedGoogle Scholar
  • 19 Pfaneuf S, Europe-Finner G N, Carrasco M P, Hamilton C H, Lopez Bernal A. Oxytocin signaling in human myometrium.  Adv Exp Med Biol. 1995;  395 453-467
    PubMedGoogle Scholar
  • 20 Pitzel L, Jarry H, Wuttke W. Demonstration of oxytocin receptors in porcine corpora lutea: effects of the cycle stage and the distribution on small and large luteal cells.  Biol Reprod. 1993;  48 640-646
    PubMedGoogle Scholar
  • 21 Sanborn B M, Dodge K, Monga M, Qian A, Wang W, Yue C. Molecular mechanisms regulating the effects of oxytocin on myometrial intracellular calcium.  Adv Exp Med Biol. 1998;  449 277-286
    PubMedGoogle Scholar
  • 22 Sapino A, Macri L, Tonda L, Bussolati G. Oxytocin enhances myoepithelial cell differentiation and proliferation in the mouse mammary gland.  Endocrinol. 1993;  133 838-842
    CrossrefPubMedGoogle Scholar
  • 23 Sernia C, Gemmell R T, Thomas W G. Oxytocin receptors in the ovine corpus luteum.  J Endocrinol. 1989;  121 117-123
    PubMedGoogle Scholar
  • 24 Sirotkin A V. Inter-relationships between nonapeptide hormones and cyclic nucleotides within cultured porcine granulosa cells.  J Endocrinol. 1996;  150 343-348
    CrossrefPubMedGoogle Scholar
  • 25 Sirotkin A V, Makarevich A V. GH regulates secretory activity and apoptosis in cultured bovine granulosa cells through the activation of the cAMP/protein kinase A system.  J Endocrinol. 1999;  163 317-327
    CrossrefPubMedGoogle Scholar
  • 26 Sirotkin A V, Makarevich A V. Growth hormone can regulate functions of porcine ovarian granulosa cells through the cAMP/protein kinase A system.  Anim Reprod Sci. 2002;  70 111-126
    PubMedGoogle Scholar
  • 27 Sirotkin A V, Makarevich A V, Kotwica J, Marnet P G, Kwon H B, Hetenyi L. Isolated porcine ovarian follicles as a model for the study of hormone and growth factor action on ovarian secretory activity.  J Endocrinol. 1998;  159 313-321
    CrossrefPubMedGoogle Scholar
  • 28 Sirotkin A V, Makarevich A V, Kwon H B, Kotwica J. Involvement of MAP kinase in the mediation of GH action on ovarian granulosa cells.  Mol Cell Endocrinol. 2003;  205 193-199
    CrossrefPubMedGoogle Scholar
  • 29 Sirotkin A V, Nitray J. The influence of oxytocin, vasopressin and their analogues on progesterone and testosterone production by porcine granulosa cells in vitro.  Ann Endocrinol. 1992;  53 32-36
    PubMedGoogle Scholar
  • 30 Sirotkin A V, Nitray J, Tarasenko L W, Uhrin P, Laurincik J, Bulla J. Effects of oxytocin, vasopressin and vasotocin and their agonists on steroid secretion by bovine granulosa cells.  Anim Reprod Sci. 1995;  39 81-87
    PubMedGoogle Scholar
  • 31 Sirotkin A V, Schaeffer H J, Mlyncek M, Missik J, Bulla J. Oxytocin affects the release of steroids, insulin-like growth factor-I, prostaglandin F2 alpha and cyclic nucleotides by human granulosa cells in vitro.  Hum Reprod. 1996;  11 152-155
    PubMedGoogle Scholar
  • 32 Skarzynski D J, Uenoyama Y, Kotwica J, Okuda K. Noradrenaline stimulates the production of prostaglandin F2 alpha in cultured bovine endometrial cells.  Biol Reprod. 1999;  60 277-282
    PubMedGoogle Scholar
  • 33 Soloff M S, Jeng Y J, Copland J A, Strakova Z, Hoare S. Signal pathways mediating oxytocin stimulation of prostaglandin synthesis in select target cells.  Exp Physiol. 2000;  85 51S-58S
    CrossrefPubMedGoogle Scholar
  • 34 Strakova Z, Copland J A, Lolait S J, Soloff M S. ERK2 mediates oxytocin-stimulated PGE2 synthesis.  Am J Physiol. 1998;  274 E634-E641
    PubMedGoogle Scholar
  • 35 Takekida S, Deguchi J, Samoto T, Matsuo H, Maruo T. Comparative analysis of the effects of gonadotropin-releasing hormone agonist on the proliferative activity, apoptosis and steroidogenesis in cultured porcine granulosa cells at varying stages of follicular growth.  Endocrine. 2000;  12 61-67
    CrossrefPubMedGoogle Scholar
  • 36 Uenoyama Y, Okuda K. Regulation of oxytocin receptors in bovine granulosa cells.  Biol Reprod. 1997;  57 569-574
    PubMedGoogle Scholar

Dr. Ph.D. A. V. Makarevich

Laboratory of Endocrinology
Research Institute of Animal Production

Hlohovska 2

94992 Nitra

Slovak Republic

Phone: + 421376546335

Fax: + 42 13 76 54 63 61

Email: makarev@vuzv.sk

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