Immature Oocyte-Specific Zap70 and Its Functional Analysis in Regulating Oocyte Maturation

  • Kim, Yun-Na (Dept. of Biomedical Science, College of Life Science, CHA University, CHA Research Institute, Fertility Center, CHA General Hospital) ;
  • Kim, Eun-Ju (Dept. of Biomedical Science, College of Life Science, CHA University, CHA Research Institute, Fertility Center, CHA General Hospital) ;
  • Kim, Eun-Young (Dept. of Biomedical Science, College of Life Science, CHA University, CHA Research Institute, Fertility Center, CHA General Hospital) ;
  • Lee, Hyun-Seo (Dept. of Biomedical Science, College of Life Science, CHA University, CHA Research Institute, Fertility Center, CHA General Hospital) ;
  • Kim, Kyeoung-Hwa (Dept. of Biomedical Science, College of Life Science, CHA University, CHA Research Institute, Fertility Center, CHA General Hospital) ;
  • Lee, Kyung-Ah (Dept. of Biomedical Science, College of Life Science, CHA University, CHA Research Institute, Fertility Center, CHA General Hospital)
  • Published : 2009.09.30

Abstract

Previously, we obtained the list of genes differentially expressed between GV and MII oocytes. Out of the list, we focused on functional analysis of Zap70 in the present study, because it has been known to be expressed only in immune cells. This is the first report about the expression and its function of Zap70 in the oocytes. Synthetic 475 bp Zap70 dsRNA was microinjected into the GV oocytes, and the oocytes were cultured in vitro. In addition to maturation rates, meiotic spindle and chromosome rearrangements, and changes in expression levels of transcripts of three kinases, Erk1/2, JNK, and p38, were determined. Zap70 is highly expressed in immature GV oocytes, and gradually decreased as oocyte matured. When dsRNA of Zap70 was injected into the GV oocytes, Zap70 mRNA specifically and completely decreased by 2 hr and its protein expression also decreased significantly. Absence of Zap70 resulted in maturation inhibition at meiosis I (57%) with abnormalities in meiotic spindle formation and chromosome rearrangement. Concurrently, mRNA expression of Erk2, JNK, and p38, were affected by Zap70 RNAi. Therefore, we concluded that Zap70 is involved in MI-MII transition by affecting expression of MAP kinases.

Keywords

References

  1. Alonso A, Rahmouni S, Williams S, van Stipdonk M, Jaroszewski L, Godzik A, Abraham RT, Schoenberger SP, Mustelin T (2003) Tyrosine phosphorylation of VHR phosphatase by ZAP-70. Nat Immunol 4:44-48. https://doi.org/10.1038/ni856
  2. Aroca P, Bottaro DP, Ishibashi T, Aaronson SA, Santos E (1995) Human dual specificity phosphatase VHR activates maturation promotion factor and triggers meiotic maturation in Xenopus oocytes. J Biol Chem 270:14229-14234. https://doi.org/10.1074/jbc.270.23.14229
  3. Burkhardt JK, Carrizosa E, Shaffer MH (2008) The actin cytoskeleton in T cell activation. Annu Rev Immunol 26:233-259. https://doi.org/10.1146/annurev.immunol.26.021607.090347
  4. Chan AC, Irving BA, Fraser JD, Weiss A (1991) The zeta chain is associated with a tyrosine kinase and upon T-cell antigen receptor stimulation associates with ZAP-70, a 70-kDa tyrosine phosphoprotein. Proc Natl Acad Sci USA 88:9166-9170. https://doi.org/10.1073/pnas.88.20.9166
  5. Chan AC, Iwashima M, Turck CW, Weiss A (1992) ZAP-70: a 70 kd protein-tyrosine kinase that associates with the TCR zeta chain. Cell 71:649-662. https://doi.org/10.1016/0092-8674(92)90598-7
  6. DeJong J (2006) Basic mechanisms for the control of germ cell gene expression. Gene 366:39-50. https://doi.org/10.1016/j.gene.2005.10.012
  7. Dombroski D, Houghtling RA, Labno CM, Precht P, Takesono A, Caplen NJ, Billadeau DD, Wange RL, Burkhardt JK, Schwartzberg PL (2005) Kinase-independent functions for Itk in TCR-induced regulation of Vav and the actin cytoskeleton. J Immunol 174:1385-1392. https://doi.org/10.4049/jimmunol.174.3.1385
  8. Fan HY, Sun QY (2004) Involvement of mitogen-activated protein kinase cascade during oocyte maturation and fertilization in mammals. Biol Reprod 70:535-547. https://doi.org/10.1095/biolreprod.103.022830
  9. Fluck M, Zurcher G, Andres AC, Ziemiecki A (1995) Molecular characterization of the murine syk protein tyrosine kinase cDNA, transcripts and protein. Biochem Biophys Res Commun 213:273-281. https://doi.org/10.1006/bbrc.1995.2126
  10. Hashimoto N, Kishimoto T (1988) Regulation of meiotic metaphase by a cytoplasmic maturation-promoting factor during mouse oocyte maturation. Dev Biol 126:242-252. https://doi.org/10.1016/0012-1606(88)90135-2
  11. Kim KH, Kim EY, Lee KA (2008) SEBOX is essential for early embryogenesis at the two-cell stage in the mouse. Biol Reprod 79:1192-1201. https://doi.org/10.1095/biolreprod.108.068478
  12. Labno CM, Lewis CM, You D, Leung DW, Takesono A, Kamberos N, Seth A, Finkelstein LD, Rosen MK, Schwartzberg PL, Burkhardt JK (2003) Itk functions to control actin polymerization at the immune synapse through localized activation of Cdc42 and WASP. Curr Biol 13:1619-1624. https://doi.org/10.1016/j.cub.2003.08.005
  13. Laurenti L, Petlickovski A, Rumi C, Gobessi S, Piccioni P, Tarnani M, Puggioni P, Marietti S, Sica S, Leone G, Efremov DG (2005) Comparison of ZAP-70/Syk mRNA levels with immunoglobulin heavy-chain gene mutation status and disease progression in chronic lymphocytic leukemia. Haematologica 90:1533-1540.
  14. Mortarino M, Gelain ME, Gioia G, Ciusani E, Bazzocchi C, Comazzi S (2009) ZAP-70 and Syk expression in canine lymphoid cells and preliminary results on leukaemia cases. Vet Immunol Immunopathol 128:395-401. https://doi.org/10.1016/j.vetimm.2008.12.010
  15. Negishi I, Motoyama N, Nakayama K, Senju S, Hatakeyama S, Zhang Q, Chan AC, Loh DY (1995) Essential role for ZAP-70 in both positive and negative selection of thymocytes. Nature 376:435-438. https://doi.org/10.1038/376435a0
  16. Roth Z, Hansen PJ (2005) Disruption of nuclear maturation and rearrangement of cytoskeletal elements in bovine oocytes exposed to heat shock during maturation. Reproduction 129:235-244. https://doi.org/10.1530/rep.1.00394
  17. Sobajima T, Aoki F, Kohmoto K (1993) Activation of mitogen-activated protein kinase during meiotic maturation in mouse oocytes. J Reprod Fertil 97:389-394. https://doi.org/10.1530/jrf.0.0970389
  18. Sun QY, Schatten H (2006) Regulation of dynamic events by microfilaments during oocyte maturation and fertilization. Reproduction 131:193-205. https://doi.org/10.1530/rep.1.00847
  19. Todd JL, Tanner KG, Denu JM (1999) Extracellular regulated kinases (ERK) 1 and ERK2 are authentic substrates for the dual-specificity protein-tyrosine phosphatase VHR. A novel role in down-regulating the ERK pathway. J Biol Chem 274:13271-13280. https://doi.org/10.1074/jbc.274.19.13271
  20. Tsuchida S, Yanagi S, Inatome R, Ding J, Hermann P, Tsujimura T, Matsui N, Yamamura H (2000) Purification of a 72-kDa protein-tyrosine kinase from rat liver and its identification as Syk: involvement of Syk in signaling events of hepatocytes. J Biochem 127:321-327. https://doi.org/10.1093/oxfordjournals.jbchem.a022610
  21. Tsujimura T, Yanagi S, Inatome R, Takano T, Ishihara I, Mitsui N, Takahashi S, Yamamura H (2001) Syk protein-tyrosine kinase is involved in neuron-like differentiation of embryonal carcinoma P19 cells. FEBS Lett 489:129-133. https://doi.org/10.1016/S0014-5793(01)02097-X
  22. Verlhac MH, Lefebvre C, Guillaud P, Rassinier P, Maro B (2000) Asymmetric division in mouse oocytes: with or without Mos. Curr Biol 10:1303-1306. https://doi.org/10.1016/S0960-9822(00)00753-3
  23. Yamada T, Fujieda S, Yanagi S, Yamamura H, Inatome R, Sunaga H, Saito H (2001) Protein-tyrosine kinase Syk expressed in human nasal fibroblasts and its effect on RANTES production. J Immunol 166:538-543. https://doi.org/10.4049/jimmunol.166.1.538
  24. Yanagi S, Inatome R, Ding J, Kitaguchi H, Tybulewicz VL, Yamamura H (2001) Syk expression in endothelial cells and their morphologic defects in embryonic Sykdeficient mice. Blood 98:2869-2871. https://doi.org/10.1182/blood.V98.9.2869
  25. Yoon SJ, Chung HM, Cha KY, Kim NH, Lee KA (2005) Identification of differential gene expression in germinal vesicle vs. metaphase II mouse oocytes by using annealing control primers. Fertil Steril 83 Suppl 1:1293-1296. https://doi.org/10.1016/j.fertnstert.2004.09.037
  26. Yoon SJ, Koo DB, Park JS, Choi KH, Han YM, Lee KA (2006) Role of cytosolic malate dehydrogenase in oocyte maturation and embryo development. Fertil Steril 86:1129-1136. https://doi.org/10.1016/j.fertnstert.2006.02.105