Cell cycle dependent regulation of gap junction coupling and apoptosis in GFSHR-17 granulosa cells
Sabrina Schlie, Karolina Mazur, Willem Bintig, Anaclet Ngezahayo
DOI: 10.4236/jbise.2010.39118   PDF    HTML     4,925 Downloads   8,559 Views   Citations


Recent results have shown that the level of gap junction coupling could modulate the induction of apoptotic reactions. We previously observed that 1H-[1,2, 4]Oxadiazole[4,3-a]quinoxalin-1-one (ODQ), a block- er of guanylyl cyclase, inhibited gap junction coupling and thereby promoted activation of characteristic apoptotic reactions such as chromatin condensation, DNA strand breaking, and formation of blebs in GFSHR-17 granulosa cells, the in vitro model for granulosa cells of the maturing ovular follicle. In the present report, we focus on the effects of ODQ with respect to the cell cycle in GFSHR-17 granulosa cells. In synchronised GFSHR-17 granulosa cells, the double whole-cell patch-clamp technique revealed that gap junction conductance in mitotic cells was reduced in comparison to cells in interphase. This reduction of gap junction conductance correlated with a reduction of non-phosphorylated Cx43 in mitotic cells. We compared the stimulation of apoptotic reactions by ODQ between cells in mitosis and in interphase. We observed that the induction of both chromatin condensation and DNA strand breaking by ODQ was increased in mitotic cells, as compared to cells in interphase. The effects of ODQ were not observed in He-La cells that do not express connexins. The results in- dicate that reduction of gap junction coupling in mitotic GFSHR-17 granulosa cells depends on phosphor- rylation of Cx43 and raises the sensitivity to stimulation of apoptosis. We propose that gap junction coupling is involved in regulation of apoptosis of granulosa cells in maturing ovular follicle.

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Schlie, S. , Mazur, K. , Bintig, W. and Ngezahayo, A. (2010) Cell cycle dependent regulation of gap junction coupling and apoptosis in GFSHR-17 granulosa cells. Journal of Biomedical Science and Engineering, 3, 884-891. doi: 10.4236/jbise.2010.39118.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Harris, A.L. (2001) Emerging issues of connexin channels: Biophysics fills the gap. Quarterly Review of Biophysics, 34(3), 325-472.
[2] White, T.W. and Paul D.L. (1999) Genetic diseases and gene knockouts reveal diverse connexin functions. Ann- ual Review of Physiology, 61(1), 283-310.
[3] Willecke, K., Eiberger, J., Degen, J., Eckardt, J.D., Ro- mualdi, A., Guldenagel, M., Deutsch, U. and Sohl, G. (2002) Structural and functional diversity of connexin genes in the mouse and human genome. The Journal of Biological Chemistry, 383(5), 725-737.
[4] Nakase, T., Fushiki, S. and Naus, C.C. (2003) Astrocytic gap junctions composed of connexin 43 reduce apoptotic neuronal damage in cerebral ischemia. Stroke, 34(8), 1987-1993.
[5] Doble, B.W., Dang, X., Ping, P., Fandrich, R.R., Nickel, B.E., Jin, Y., Cattini, P.A. and Kardami, E. (2003) Phosphorylation of serine 262 in the gap junction protein connexin-43 regulates DNA synthesis in cell-cell contact forming cardiomyocytes. Journal of Cell Science, 117(3), 507-514.
[6] Gorbe, A., Becker, D.L., Dux, L., Stelkovics, E., Krenacs, L., Bagdi, E. and Krenacs, T. (2005) Transient upregulation of connexin43 gap junctions and synchronized cell cycle control precede myoblast fusion in regenerating skeletal muscle in vivo. Histochemistry and Cell Biology, 123(6), 573-583.
[7] Gorbe, A., Krenacs, T., Cook, J E. and Becker, D.L. (2007) Myoblast proliferation and syncytial fusion both depend on connexin43 function in transfected skeletal muscle primary cultures. Experimental Cell Research, 313(6), 1135-1148.
[8] Zhang, Y.W., Morita, I., Ikeda. M., Ma, K.W. and Mu- rota, S. (2001) Connexin43 suppresses proliferation of osteosarcoma U2OS cells through post-transcriptional regulation of p27. Oncogene, 20(31), 4138-4149.
[9] Zhang, Y.W., Chen, X., Wu, D., Liu, W., Wang, J., Feng, Z., Cai, G., Fu, B., Hong, Q. and Du, J. (2006) Downregulation of connexin 43 expression by high glucose induces senescence in glomerular mesangial cells. Journal of the American Society of Nephrology, 17(6), 1532-1542.
[10] Ngezahayo, A., Altmann, B., Steffens, M. and Kolb, H. A. (2005) Gap junction coupling and apoptosis in GF- SHR-17 granulosa cells. Journal of Membrane Biology 204, 137-144.
[11] Solan, J.L., Fry, M.D., TenBroek, E.M. and Lampe, P.D. (2003) Connexin43 phosphorylation at S368 is acute during S and G2/M and in response to protein kinase C activation. Journal of Cell Science, 116(pt11), 2203- 2211.
[12] Keren, T.I., Dantes, A., Sprengel, R. and Amsterdam, A. (1993) Establishment of steroidegenic granulosa cell li- nes expressing follicle stimulating hormone receptors. Molecular and Cellular Endocrinology, 95, R1-R10.
[13] Okuma, A., Kuraoka, A., Iida, H., Inai, T., Wasano, K. and Shibata, Y. (1996) Colocalization of connexin 43 and connexin 45 but absence of connexin 40 in granulosa cell gap junctions of rat ovary. Journal Reprod Fertil, 107(2), 255-264.
[14] Ackert, C.L., Gittens, J.E.I., O’Brien, M.J., Eppig, J.J. and Kidder, G.M. (2001) Intercellular communication via connexin43 gap junctions is required for ovarian folicu- logenesis in the mouse. Developmental Biology, 233(2), 248-270.
[15] Wiesen, J.F. and Midgley, R.A. (1994) Expression of connexin43 gap junction messenger ribonucleic acid and protein during follicular atresia. Biology of Reproduction, 50(2), 336-348.
[16] Wright, C.S., Becker, D.L., Lin, J.S., Warner, A.E. and Hardy, K. (2001) Stage-specific and differential express- ion of gap junctions in the mouse ovary: Connexin-spe- cific roles in follicular regulation. Reproduction, 121, 77- 88.
[17] Quirk, M., Cowan, R.G. and Harman, R.M. (2004) Prog- esterone receptor and the cell cycle modulate apoptosis in granulosa cells. Endocrinol, 145(11), 5033-5043.
[18] Sasson, R. and Amsterdam, A. (2003) Pleiotropic anti- apoptotic activity of glucocorticoids in ovarian follicular cells. Biochemical Pharmacology, 66(8), 1393-401.
[19] Lampe, P.D. and Lau, A.F. (2004) The effects of connexin phosphorylation on gap junctional communication. The International Journal of Biochemistry & Cell Biology, 36(7), 1171-1186.
[20] Ngezahayo, A., Altmann, B. and Kolb, H.A. (2003) Regulation of gap junctional coupling, ion fluxes and cell volume by cGMP in GFSHR-17 granulosa cells. Journal of Membrane Biology, 194(3), 165-176.
[21] Van Rossum, G.S.A.T., Vlug, A.S., van den Bosch, H., Verkleij, A.J. and Boonstra, J. (2001) Cytosolic phosph- olipase A2 activity during the ongoing cell cycle. Journal of Cellular Physiology, 188, 321-328.
[22] Schumer, S.T. and Cannistra, S.A. (2003) Granulosa cell tumor of the ovary. Journal of Clinical Oncology, 21(6), 1180-1189.
[23] Barker, C.J., Wright, J., Hughes, P.J., Kirk, C.J. and Michel, R.H. (2004) Complex changes in cellular inositol phosphate complement accompany transit through the cell cycle. Biochemical Journal, 380(pt 2), 465-473.
[24] Musil, L.S., Cunningham, B.A., Edelmann, G.M. and Goodenough, D.A. (1990) Differential phosphorylation of the gap junction protein connexin43 in junctional communication-competent and deficient cell lines. The Journal of Cell Biology, 111(5 pt 1), 2077-2088.
[25] Bittman, K.S. and LoTurco, J.J. (1999) Differential regulation of connexin 26 and 43 in murine neocortical precursors. Cerebral cortex, 9(2), 188-195.
[26] Solan, J.L. and Lampe, P.D. (2007) Key connexin43 phosphorylation events regulate the gap junction life cycle. Journal of Membrane Biology, 217(1-3), 35-41.
[27] Jin, E.J., Lee, S.Y., Jung, J.C., Bang, O.S. and Kang, S.S. (2008) TGF-β3 inhibits chondrogenesis of cultured chick leg bud mesenchymal cells via downregulation of connexin 43 and integrin β4. Journal of Cellular Physiology, 214, 345-353.
[28] Becker, D.L., Webb, K.F., Thrasivoulou, C., Lin, C.C., Nadershahi, R., Tsakiri, N. and Cook, J.E. (2007) Multiphoton imaging of chick retinal development in relation to gap junctional communication. The Journal of Physiology, 585(pt 3), 711-719
[29] Pearson, R.A., Lüneborg, N.L., Becker, D.L. and Mobbs, P. (2005) Gap junctions modulate interkinetic nuclear movement in retinal progenitor cells. The Journal of Neuroscience, 25(46), 10803-10814.
[30] Andreu-Vieyra, C., Chen, R. and Matzuk, M.M. (2007) Effects of granulosa cell-specific deletion of Rb in Inha-α null female mice. Endocrinology, 148(8), 3837-3849.

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