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Preantral Follicle Growth is Regulated by c-Jun-N-Terminal Kinase (JNK) Pathway

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Abstract

c-Jun N-terminal kinase (JNK) pathway has been shown to be essential for cell cycle progression and mitosis. We previously showed that this pathway is activated in mitotic granulosa cells of follicles from transitional to antral stages. In this study, we, therefore, aimed to investigate whether this signaling pathway has any effect on in-vitro growth of murine preantral follicles and granulosa cell cycle control. Two structurally different pharmacologic JNK inhibitors, SP600125 and AS601245, were used in the experiments. First their inhibitory concentrations were determined in granulosa cells by Western blot analysis. Then preantral follicles isolated from immature and adult C57BL/6 mice were cultured in matrigel and standard culture plates for 6 days with these inhibitors. Spontaneously immortalized rat granulosa cells (SIGCs) were first synchronized at G1/S and G2/M stages of cell cycle and then treated with JNK inhibitors. Cell cycle progression was analyzed with Bromodeoxyuridine (BrdU) assay and flow cytometry analysis. Both inhibitors significantly inhibited phosphorylation of c-Jun in granulosa cells at 25, 50, and 100 μmol/L concentrations. Isolated preantral follicles cultured with these inhibitors exhibited arrested growth in culture in a dose-dependent manner. Cell cycle analyses showed that both inhibitors impair the progression of cell cycle at S phase and G2/M transition of granulosa cells. These results suggest that JNK pathway is essential for in vitro growth of preantral follicle growth and regulates both S phase and G2/M stages of cell cycle in granulosa cells.

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References

  1. Weston CR, Davis RJ. The JNK signal transduction pathway. Curr Opin Cell Biol. 2007;19(2):142–149.

    Article  CAS  Google Scholar 

  2. Johnson RS, van Lingen B, Papaioannou VE, Spiegelman BM. A null mutation at the c-jun locus causes embryonic lethality and retarded cell growth in culture. Genes Dev. 1993;7(7B):1309–1317.

    Article  CAS  Google Scholar 

  3. Oktay K, Oktay M. Immunohistochemical analysis of tyrosine phosphorylation and AP-1 transcription factors c-Jun, Jun D, and Fos family during early ovarian follicle development in the mouse. Appl Immunohistochem Mol Morphol. 2004;12(4):364–369.

    Article  CAS  Google Scholar 

  4. Gaillard P, Jeanclaude-Etter I, Ardissone V, et al. Design and synthesis of the first generation of novel potent, selective, and in vivo active (benzothiazol-2-yl)acetonitrile inhibitors of the c-Jun N-terminal kinase. J Med Chem. 2005;48(14):4596–4607.

    Article  CAS  Google Scholar 

  5. Oktay K, Buyuk E, Oktem O, Oktay M, Giancotti FG. Jun amino terminal kinase regulates G2/M transition through a mechanism upstream of aurora-B. Cell Cycle. 2008;7(4):533–541.

    Article  CAS  Google Scholar 

  6. Stein LS, Stoica G, Tilley R, Burghardt RC. Rat ovarian granulosa cell culture: a model system for the study of cell-cell communication during multistep transformation. Cancer Res. 1991;51(2):696–706.

    PubMed  CAS  Google Scholar 

  7. Kleinman HK, Martin GR. Matrigel: basement membrane matrix with biological activity. Semin Cancer Biol. 2005;15(5):378–386.

    Article  CAS  Google Scholar 

  8. Oktem O, Oktay K. The role of extracellular matrix and activin-A in in-vitro growth and survival of murine preantral follicles. Reprod Sci. 2007;14(4):358–366.

    Article  CAS  Google Scholar 

  9. Oktay K, Buyuk E, Oktem O. Deciphering Early Folliculogenesis: Jun Amino Terminal Kinase (JNK) plays a key role in preantral follicle growth. Fertil Steril. 2003;80(3):26–27.

    Article  Google Scholar 

  10. Oktem O, Oktay K. Role of extracellular matrix, Activin-A and mitogen activated protein kinase (MAPK) signaling pathways in preantral follicle growth and survival in the mouse. Fertil Steril. 2005;84(S1):384–385.

    Article  Google Scholar 

  11. Oktay K, Karlikaya G, Ojakian GK, Oktay M. Interaction of extracellular matrix and activin-A in the initiation of follicle growth in the mouse ovary. Biol Reprod. 2000;63(2):457–461.

    Article  CAS  Google Scholar 

  12. Oktay M, Wary KK, Dans M, Birge RB, Giancotti FG. Integrin-mediated activation of focal adhesion kinase is required for signaling to Jun NH2-terminal kinase and progression through the G1 phase of the cell cycle. J Cell Biol. 1999;145(7):1461–1469.

    Article  CAS  Google Scholar 

  13. Angel P, Karin M. The role of Jun, Fos and the AP-1 complex in cell-proliferation and transformation. Biochim Biophys Acta. 1991;1072(2‐3):129–157.

    PubMed  CAS  Google Scholar 

  14. Davis RJ. Signal transduction by the JNK group of MAP kinases. Cell. 2000;103(2):239–252.

    Article  CAS  Google Scholar 

  15. Kovary K, Bravo R. The jun and fos protein families are both required for cell cycle progression in fibroblasts. Mol Cell Biol. 1991;11(9):4466–4472.

    Article  CAS  Google Scholar 

  16. Riabowol KT, Vosatka RJ, Ziff EB, Lamb NJ, Feramisco JR. Microinjection of fos-specific antibodies blocks DNA synthesis in fibroblast cells. Mol Cell Biol. 1988;8(4):1670–1676.

    Article  CAS  Google Scholar 

  17. Yamamoto K, Ichijo H, Korsmeyer SJ. BCL-2 is phosphorylated and inactivated by an ASK1/Jun N-terminal protein kinase pathway normally activated at G2/M. Mol Cell Biol. 1999;19(12):8469–8478.

    Article  CAS  Google Scholar 

  18. Kim JA, Lee J, Margolis RL, Fotedar R. SP600125 suppresses Cdk1 and induces endoreplication directly from G2 phase, independent of JNK inhibition. Oncogene. 2010;29(11):1702–1716.

    Article  CAS  Google Scholar 

  19. Sriraman V, Modi SR, Bodenburg Y, Denner LA, Urban RJ. Identification of ERK and JNK as signaling mediators on protein kinase C activation in cultured granulosa cells. Mol Cell Endocrinol. 2008;294(1–2):52–60.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Laboratory of Fertility Preservation and Molecular Reproduction, Departments of Obstetrics & Gynecology, Cell Biology & Anatomy, and Medicine, New York Medical College, Valhalla, New York, 10595, USA

    Ozgur Oktem MD, Erkan Buyuk MD &  Kutluk Oktay MD

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  1. Ozgur Oktem MD
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  2. Erkan Buyuk MD
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  3. Kutluk Oktay MD
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Correspondence to Kutluk Oktay MD.

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Oktem, O., Buyuk, E. & Oktay, K. Preantral Follicle Growth is Regulated by c-Jun-N-Terminal Kinase (JNK) Pathway. Reprod. Sci. 18, 269–276 (2011). https://doi.org/10.1177/1933719110385709

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