Hostname: page-component-586b7cd67f-tf8b9 Total loading time: 0 Render date: 2024-11-26T15:56:52.801Z Has data issue: false hasContentIssue false
  • English
  • Français

Co-culture with pig membrana granulosa cells modulates the activity of cdc2 and MAP kinase in maturing cattle oocytes

Published online by Cambridge University Press:  26 September 2008

Jan Motlík ,
Peter Šutovský ,
Jaroslav Kalous ,
Michal Kubelka ,
Jiří Moos  and
Richard M. Schultz
Jan Motlík*
Affiliation:
Insitute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Liběchov, Czech Republic.
Peter Šutovský
Affiliation:
Insitute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Liběchov, Czech Republic.
Jaroslav Kalous
Affiliation:
Insitute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Liběchov, Czech Republic.
Michal Kubelka
Affiliation:
Insitute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Liběchov, Czech Republic.
Jiří Moos
Affiliation:
Insitute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Liběchov, Czech Republic.
Richard M. Schultz
Affiliation:
Insitute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Liběchov, Czech Republic.
*
Dr Jan Motlík, Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, 277 21 Liběchov, Czech Republic. Telephone: +42 206 697147. Fax: +42 206 697186. e-mail: [email protected].
Get access Rights & Permissions [Opens in a new window]

Summary

Bovine cumulus-enclosed oocytes, initially cultured up to diakinesis (8h of initial culture) or metaphase I (12h of initial culture), were subsequently co-cultured for 6 h in contact with pig membrana granulosa (PMG) cells and then assayed for histone H1 and MAP kinase activities. In addition, the phosphorylation state of ERK 1,2 proteins was determined by Western blotting. The alterations in nuclear envelope breakdown, meiotic spindle formation and the patterns of chromosome condensation were analysed by immunofluorescence and transmission electron microscopy. The diakinesis-stage oocytes (initially cultured for 8h) already possessed high histone H1 kinase and MAP kinase activities that were correlated with condensed and partially individualised chromosomes. The ERK 1 and most ERK 2 proteins were partly phosphorylated. Following the 6h co-culture of these oocytes with PMG a rapid decrease in MAP kinase activity and a slower decrease in histone H1 kinase occurred, as well as ERK 1 and ERK 2 dephosphorylation. Both kinase activities and ERK 1,2 phosphorylation were fully restored following the release of the oocytes from co-culture and a subsequent culture in the absence of PMG. Moreover, the clumped bivalents were reindividualised and 56% of these oocytes reached metaphase II after 20 h of culture without PMG. The metaphase I oocytes, initially cultured for 12 h, displayed a fusiform meiotic spindle and a metaphase array of chromosomal bivalents, accompanied by high levels of both histone H1 and MAP kinase activity. Co-culture of MI oocytes with PMG abolished the activity of both kinases and caused the dephosphorylation of ERK 1 and ERK 2. Furthermore, the spindle microtubules were depolymerised and the chromosomal bivalents clumped into a single mass. Neither of the protein kinase activities nor the meiotic spindle were restored following subsequent culture in the absence of PMG for up to 20 h. These observations indicate that under in vitro conditions membrana granulosa cells can cause a prompt decrease in histone H1 and MAP kinase activities, and metaphase I oocytes. While these events are fully reversible in late diakinesis oocytes, metaphase I oocytes did not complete maturation after release from co-culture.

Keywords

Porcine granulosa cellsCattle oocytesCo-cultureMeiosisMAP and H1 kinase
Type
Article
Information
Zygote , Volume 4 , Issue 3 , August 1996 , pp. 247 - 256
Copyright
Copyright © Cambridge University Press 1996

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Buendia, B., Draetta, G. & Karsenti, E.. (1992). Regulation of microtubule nucleating activity of centrosomes in Xenopus egg extracts. J. Cell Biol 116, 1431–42.CrossRefGoogle ScholarPubMed
Byskov, A.G., Anderson, C.Y., Nordholm, L., Thogersen, H., Guoliang, X., Wassmann, O., Andersen, J.V., Guddal, E..& Roed, T.. (1995). Chemical structure of sterols that activate oocyte meiosis. Nature 374, 559–62.CrossRefGoogle ScholarPubMed
Calarco, P.G., Donahue, R.P. & Szollosi, D.. (1972). Germinal vesicle breakdown in the mouse oocyte. J. Cell Sci. 10, 369–85.CrossRefGoogle ScholarPubMed
Centonze, V.E. & Borisy, G.G.. (1990). Nucleation of micro-tubules from mitotic centrosomes is modulated by a phosphorylated epitope. J. Cell Sci. 95, 405–11.CrossRefGoogle Scholar
Chesnel, F. & Eppig, J.J.. (1995). Induction of precocious germinal vesicle breakdown (GVB) by GVB-incompetent mouse oocytes: possible role of mitogen-activated protein kinases rather than p34cdc2 kinase. Biol. Reprod. 52, 895–902.CrossRefGoogle ScholarPubMed
Colona, R., Cecconi, S., Tatone, C., Mangia, F., & Buccione, R.. (1989). Somatic cell–oocyte interactions in mouse oogenesis: stage specific regulation of mouse oocyte protein phosphorylation by granulosa cells. Dev. Biol 133, 305–8.CrossRefGoogle Scholar
De Loos, F.A.M., Zeinstra, E. & Bevers, M.M.. (1994). Follicular wall maintains meiotic arrest in bovine oocytes cultured in vitro. Mol. Reprod. Dev. 39, 162–5.CrossRefGoogle ScholarPubMed
De Pennart, H., Houliston, E. & Maro, B.. (1988). Post-translational modifications of tubulin and the dynamics of microtubules in mouse oocytes and zygotes. Biol. Cell 64, 375–8.Google ScholarPubMed
Dessev, C., Ivcheva-Dessev, C.., Bischoff, J., Beach, D. & Goldman, R.. (1991). A complex containing p34cdc2 and cyclin B phosphorylates the nuclear lamin and disassembles the nuclei of clam oocytes in vitro. J. Cell Biol 112, 523–33.CrossRefGoogle ScholarPubMed
Downs, S.M. & Eppig, J.J.. (1987). Induction of mouse oocyte maturation in vivo by perturbants of purine metabolism. Biol. Reprod 36, 431–7.CrossRefGoogle ScholarPubMed
Fesquet, D., Labbé, J.C., Derancourt, J., Capony, J.P., Galas, S., Girard, F., Lorca, T., Shuttleworth, J., Dorée, M. & Cavadore, J.C.. (1993). The MO15 gene encodes the catalytic subunit of a protein kinase that activates cdc2 and other cyclin-dependent kinases (CDKs) through phosphorylation of Thr161 and its homologues. EMBO J. 12, 3111–21.CrossRefGoogle ScholarPubMed
Gavin, A.C., Cavadore, J.C. & Schorderet-Slatkine, S.. (1994). Histone H1 kinase activity, germinal vesicle breakdown and M phase entry in mouse oocytes. J. Cell Sci. 107, 275–83.CrossRefGoogle Scholar
Gotoh, Y., Moriyama, K., Matsuda, S., Okumura, E., Kishimoto, T., Kawasaki, H., Suzuki, K., Yahara, I., Sakai, H. & Nishida, E.. (1991). Xenopus M phase MAP kinase: isolation of its cDNA and activation by MPF. EMBO. J. 10, 2661–8.CrossRefGoogle ScholarPubMed
Guoliang, X., Byskov, A.G. & Andersen, C.Y.. (1994). Cumulus cells secrete a meiosis-inducing substance by stimulation with forskolin and dibutyric cyclic adenosine monophosphate. Mol. Reprod. Dev. 39, 17–24.CrossRefGoogle Scholar
Haccard, O., Lewellyn, A., Hartley, R.S., Erikson, E..& Maller, J.L.. (1995). Induction of Xenopus oocyte maturation by MAP kinase. Dev. Biol 168, 677–82.CrossRefGoogle ScholarPubMed
Harrouk, W. & Clarke, H.J.. (1993). Sperm chromatin acquires an activity that induces microtubule assembly during residence in the cytoplasm of metaphase oocytes of the mouse. Chromosoma 102, 279–86.CrossRefGoogle ScholarPubMed
Inoue, M., Naito, K., Aoki, F., Toyoda, Y. & Sato, E.. (1995). Activation of mitogen-activated protein kinase during meiotic maturation in porcine oocytes. Zygote 3, 265–71.CrossRefGoogle ScholarPubMed
Kaláb, P., Kubiak, J.Z., Verlhac, M.H., Colledge, W.H. & Maro, B.. (1996). Activation of p9Orsk during meiotic maturation and first mitosis in mouse oocytes and eggs: MAP kinase-independent and -dependent activation. Development. (in press).CrossRefGoogle ScholarPubMed
Kalous, J., Sutovský, P., Rimkevičová, Z., Shioya, Y., Lie, B.L. & Motlík, J.. (1993). Pig membrana granulosa cells prevent resumption of meiosis in cattle oocytes. Mol. Reprod. Dev. 34, 58–64.CrossRefGoogle ScholarPubMed
Kubelka, M., Rimkevičová, Z., Guerrier, P. & Motlík, J.. (1995). Inhibition of protein synthesis affects histone H1 kinase, but not chromosome condensation activity, during the first meiotic division of pig oocytes. Mol. Reprod. Dev. 41, 63–9.CrossRefGoogle Scholar
Laemmli, U.K.. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 11713–20.CrossRefGoogle ScholarPubMed
Langan, T.A., Gautier, J., Lohka, M., Hollingsworth, R., Moreno, P.N., Nurse, P.N., Maller, J.L. & Sclafani, R.. (1989). Mammalian growth-associated H1 histone kinase: a homolog of cdc2+/CDC28 protein kinase controlling mitotic entry in yeast and frog cells. Mol. Cell. Biol 9, 3860–8.Google ScholarPubMed
Lohka, M.J., Hayes, M. & Maller, J.. (1988). Purification of maturation-promoting-factor, an intracellular regulator of early mitosis events. Proc. Natl. Acad. Sci. USA 85, 3009–13.CrossRefGoogle Scholar
Motlík, J.. (1989). Cytoplasmic aspects of oocyte growth and maturation in mammals. J. Reprod. Fertil 38, 17–25.Google ScholarPubMed
Motlík, J. & Kubelka, M.. (1990). Cell-cycle aspects of growth and maturation of mammalian oocytes. Mol. Reprod. Dev. 27, 366–75.CrossRefGoogle ScholarPubMed
Moos, J., Visconti, P.E., Moor, G.D., Schultz, R.M. & Kopf, G.S.. (1995). Potential role of mitogen-activated protein kinase in pronuclear envelope assembly and disassembly following fertilization of mouse eggs. Biol. Reprod. 53, 692–9.CrossRefGoogle ScholarPubMed
Nagyové, E., Kalous, J., Šutovský, P. & Motlík, J.. (1993). Is cAMP decrease essential for resumption of meiosis in mouse oocytes?. Reprod. Nutr. Dev. 33, 419–28.CrossRefGoogle Scholar
Parker, L.L., Atherton-Fessler, S. & Piwnica-Worms, H.. (1992). Inactivation of the p34cdc2–cyclin B complex by the human weel tyrosine kinase. Science 257, 1955–7.CrossRefGoogle Scholar
Parker, L.L., Walter, S.A., Young, P.G. & Piwnica-Worms, H. (1993). Phosphorylation and inactivation of the mitotic inhibitor Wel by the nim1/cdrl kinase. Nature 363, 736–8.CrossRefGoogle Scholar
Peter, M., Sanghera, J.S., Pelech, S.L. & Nigg, E.A.. (1992). Mitogen-activated protein kinases phosphorylate nuclear lamins and display sequence specificity overlapping that of mitotic protein kinase P34cdc2. Eur. J. Biochem. 205, 287–94.CrossRefGoogle ScholarPubMed
Pincus, G. & Enzmann, E.V.. (1935). The comparative behavior of mammalian eggs in vivo and in vitro. J. Exp. Med. 62, 665–75.CrossRefGoogle ScholarPubMed
Russell, P. & Nurse, P.. (1987). Negative regulation of mitosis by wee1+ functions in a regulatory network of protein kinase homologs controlling the initiation of meiosis. Cell 49, 569–76.CrossRefGoogle Scholar
Schatten, G., Simerly, C., Asai, D.J., Szoke, E., Cooke, P. & Schatten, H.. (1988). Acetylated alpha-tubulin in micro-tubules during mouse fertilization and early development. Dev. Biol 130, 74–86.CrossRefGoogle Scholar
Schultz, R.M.. (1990). Meiotic maturation of mammalian oocytes. In The Biology and Chemistry of Fertilization, ed. Wassarman, P.M., 1, 77–104. Boca Raton, Fl: CRC Press.Google Scholar
Shibuya, E.K., Boulton, T.G., Cobb, M.H. & Ruderman, J.V.. (1992). Activation of p42 MAP kinas and the release of oocytes from cell cycle arrest. EMBO J. 111, 3963–75.CrossRefGoogle Scholar
Sirard, M.A. & First, N.L.. (1988). In vitro inhibition of oocyte nuclear maturation in the bovine. Biol. Reprod 39, 229–34.CrossRefGoogle ScholarPubMed
Solomon, M.J., Lee, T. & Kirschner, M.W.. (1992). Role of phosphorylation in p32cdc2 activation: identification of an activating kinase. Mol. Biol. Cell 3, 13–27.CrossRefGoogle Scholar
Solomon, M.J., Harper, J.W. & Shuttleworth, J.. (1993). CAK, the p34cdc2 activating kinase, contains a protein identical or closely related to P40MO15. EMBO J. 12, 3133–42.CrossRefGoogle ScholarPubMed
Šutovský, P., Fléchon, J.E., Fléchon, B., Motlík, J., Peynot, N., Chesne, P. & Heyman, Y.. (1993). Dynamic changes of gap junctions and cytoskeletons during in vitro culture of cattle oocyte cumulus complexes. Biol. Reprod 49, 1277–87.CrossRefGoogle ScholarPubMed
Verde, F., Labbé, J.C., Dorrée, M. & Karsenti, E.. (1990). Regulation of microtubule dynamics by cdc2 kinase in cell-free extracts of Xenopus eggs. Nature 343, 233–8.CrossRefGoogle ScholarPubMed
Verlhac, M.H., De Pennart, H., Maro, B., Cobb, M.H. & Clarke, H.J.. (1993). MAP kinase becomes stably activated at metaphase and is associated with microtubule-organizing centers during meiotic maturation of mouse oocytes. Dev. Biol 158, 330–40.CrossRefGoogle ScholarPubMed
Verlhac, M.H., Kubiak, J.Z., Clarke, H.J. & Maro, B.. (1994). Microtubule and chromatin behavior follow MAP kinase activity but not MPF activity during meiosis in mouse oocytes. Development 120, 1017–25.CrossRefGoogle Scholar
Viklický, V., Dráber, P., Hašek, J. & Bártek, J.. (1982). Production and characterization of monoclonal antitubulin antibody. Cell Biol. Int. Rep. 6, 725–33.CrossRefGoogle ScholarPubMed