표제지
목차
국문 초록 7
ABSTRACT 9
I. 서론 11
1. 세포주기와 세포검사지점 11
2. Spindle assembly checkpoint (SAC) 13
3. 난자의 세포주기 15
4. 세포주기 조절 약물 19
II. 목적 22
III. 연구재료 및 방법 23
1. 생쥐 난자세포 준비 23
2. MK1775 처리 및 난자 성숙 유도 23
3. Chromosome spreading 24
4. 면역세포화학법(Immunocytochemistry, ICC) 25
5. 인산화 효소 활성 분석법(kinase activity assay) 26
6. Western blot 26
IV. 결과 27
1. Wee1B 저해에 의한 난자 세포주기 변화 27
2. MK1775 처리에 의한 세포주기 정지 27
3. Wee1B 저해에 의한 MPF 활성 38
4. MI-MII 전환 중 Wee1B 조절 40
V. 고찰 50
VI. 결론 54
VII. 참고문헌 55
Figure 1. Cell cycle 12
Figure 2. Spindle assembly checkpoint 14
Figure 3. Meiosis 17
Figure 4. MPF activity during oocyte maturation 18
Figure 5. Regulation of Wee1B activity. 21
Figure 6. Inhibition of Wee1B causes early GVBD and blocks the polar body extrusion. 28
Figure 7. Inhibition of Wee1B induces the arrest of oocyte at metaphase I. 32
Figure 8. MK1775 treatment after GVBD also inhibits PB extrusion. 35
Figure 9. The removal of MK1775 resumed PB extrusion of oocytes 37
Figure 10. MPF activity does not decrease at MI-MII transition in MK1775 treated oocytes. 39
Figure 11. Experimental Scheme to elucidate Wee1B regulation pathway. 41
Figure 12. Effect of various kinase regulation agents on the MK1775 treated oocyte. 42
Figure 13. Forskolin, A23187 or PD98059 do not release metaphase I arrest of MK1775 treated oocyte. 45
Figure 14. Inhibition of Aurora kinases causes spindles organization defects in MK1775 treated oocyte. 46
Figure 15. Dinaciclib induces metaphase I exit and nucleation of Wee1B inhibited oocyte. 48
Figure 16. Reversine leads to polar body extrusion and sister chromatid separation of MK1775 treated oocyte. 49