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MAPK Activity in Porcine Oocytes Maturing InVitro

유사분열 활성화 단백질 효소가 돼지난자의 체외성숙에 미치는 영향

  • 이재달 (혜전대학 애완동물관리과)
  • Received : 2010.03.25
  • Accepted : 2010.06.18
  • Published : 2010.06.30

Abstract

In this study, we determined effects of the mitogen-activated protein kinase (MAPK) inhibitor, U0126 on meiotic maturation, microtubule organization and actin filament assembly in the porcine oocyte. The phosphorylated MAPK was first detected at 12 h after the initiation of maturation cultures, fully activated at 24h, and remained until metaphase II. Treatment of germinal vesicle (GV) stage oocytes with $20{\mu}M$ U0126 completely blocked MAPK phosphorylation, but germinal vesicle breakdown (GVBD) was normally proceeded. However, the oocytes didn‘t progress to the metaphase I. The inhibition of MAPK resulted in abnormal spindles. In oocytes treated with U0126 after GVBD, polar body extrusion was normal, but the organization of the metaphase plate and chromosome segregation were abnormal. In conclusion, MAPK activity plays an important regulatory role in GV chromatin configuration and meiotic progress in porcine oocyte maturation.

본 연구에서는 MAPK 저해제인 U0126이 난자성숙과정에서 특히 감수분열, 미세소관 형성 그리고 액틴 필 라먼트 형성에 미치는 영향을 조사하였다. 그 결과 MAPK 단백질은 12시간째에 인산화되기 시작하여, 24시간째에 대부분 인산화 되었고 metaphase II에 이르기 까지 유지되었다. 배포단계(GV)에 있는 난자를 U0126의 $20{\mu}M$ 농도로 처리하였을 때 MAPK의 인산화가 완전히 억제되었으나 배포의 파열 단계(GVBD)로의 성숙에는 진행하였으나, metaphase I까지는 발달하지 못하였다. 또한 MAPK 저해제로 인해 비정상적인 방추사의 형성을 초래하였다. 난자를 배포의 파열단계(GVBD) 이후에 U0126을 처리하였을 때 극체의 방출은 정상 이였으나 중기 판의 배열과 염색체의 분열은 비정상적 이였다. 결론적으로, 유사분열 활성화 효소단백질인 MAPK의 활성은 돼지 난자의 체외성숙과정에서 배포단계(GV)의 염색체의 배열과 감수분열의 완성에 중요한 조절 인자임을 이번 연구를 통해 알 수 있었다.

Keywords

References

  1. Sun QY, Schatten H. Regulation of dynamic events by microfilaments during oocyte maturation and fertilization. Repro -duction 131193-205. 2006.
  2. Sun QY, Lai L, Park KW, Kuhholzer B, Prather RS, Schatten H. Dynamic events are differently mediated by microfilaments, microtubules, and mitogen-activated protein kinase during porcine oocyte maturation and fertilization invitro. Biol Reprod 64871- 889. 2001 https://doi.org/10.1095/biolreprod64.3.871
  3. Favata MF, Horiuchi KY, Manos E J. Identification of a novel inhibitor of mitogen-activated protein kinase kinase. J Biol Chem 273 18623-32. 1998. https://doi.org/10.1074/jbc.273.29.18623
  4. Lee J, Miyano T, Moor RM. Localization of phosphorylated MAP kinase during the transition from meiosis I to meiosis II in pig oocytes. Zygote 8119-25. 2000. https://doi.org/10.1017/S0967199400000897
  5. Tatemoto H, Muto N. Mitogen-activated protein kinase regulates normal transition from metaphase to interphase following parthenogenetic activation in porcine oocytes. Zygote 9 15-23. 2001. https://doi.org/10.1017/S0967199401001034
  6. Pelech SL, Sanghera JS. MAP kinases: charting the regulatory pathways. Science 257 1355-6. 1992. https://doi.org/10.1126/science.1382311
  7. Davis R. The mitogen-activated protein kinase signal transduction pathway. J Biol Chem 268 14553-14556. 1993.
  8. Morishima-Kawashima M, Kosik KS. The pool of MAP kinase associated with microtubules in small but constitutively active. Mol Biol Cell 7893-905. 1996. https://doi.org/10.1091/mbc.7.6.893
  9. Inoue M, Naito K, Nakayama T, Sato E. Mitogen-activated protein kinase translo -cates into the germinal vesicle and induces germinal vesicle breakdown in porcine oocytes. Biol Reprod 58 130-136. 1998. https://doi.org/10.1095/biolreprod58.1.130
  10. Verlhac MH, Pennart HD, Maro B, Cobb MH, Clarke HJ. MAP kinase becomes stably activated at metaphase and in associated with microtubuleorganizing centers during meiotic maturation of mouse oocytes. Dev Biol 158 330-340. 1993. https://doi.org/10.1006/dbio.1993.1192
  11. Araki K, Naito K, Haraguchi S, Suzuki R, Yokoyama M, Inoue M, Aizawa S, Toyoda Y, Sato E. Meiotic abnormalities of c-mos knockout mouse oocytes: activation after first meiosis or entrance into third meiotic metaphase. Biol Reprod 551 315-24. 1996.
  12. Choi T, Fukasawa K, Zhou R, Tessarollo L, Borror K, Resau J, Vande Woude GF. The Mos/mitogen -activated protein kinase (MAPK) pathway regulates the size and degradation of the first polar body in maturing mouse oocytes. Proc Natl Acad Sci USA 93 7032-7035. 1996. https://doi.org/10.1073/pnas.93.14.7032