CAJ | 학술논문

为了阐明p38MAPK（p38丝裂原活化蛋白激酶）的激活在斑马鱼（Danio rerio）卵母细胞第一次减数分裂恢复中的作用，用卵泡刺激素（ FSH）、 SB203580（ p38MAPK 抑制剂）、 PKA 激活剂 forskolin 单独作用及 FSH 与SB203580、 FSH与PKA抑制剂H89共同培养斑马鱼卵母细胞。采集不同培养时间的卵母细胞，用SDS-PAGE电泳和Western Blot蛋白质免疫印迹技术检测p38MAPK磷酸化，观察生发泡破裂（GVBD）情况。结果表明，斑马鱼卵母细胞在FSH诱发下， p38 MAPK活性明显升高；单独添加SB203580斑马鱼卵母细胞p38 MAPK活性受到明显抑制，同一时间GVBD发生率均低于对照组以及同时添加FSH与SB203580的实验组； FSH与SB203580共同作用组斑马鱼卵母细胞， GVBD发生率与SB203580实验组差异性不显著，与FSH组相比显著降低。用H89与FSH共同培养卵母细胞时，虽然p38MAPK仍被激活但量很少。用forskolin单独培养卵母细胞， p38MAPK能被激活，且比对照组明显升高。结果提示，在FSH诱导的斑马鱼卵母细胞减数分裂恢复过程中p38MAPK被激活，且p38MAPK的激活是PKA依赖性的。
위료천명p38MAPK（p38사렬원활화단백격매）적격활재반마어（Danio rerio）란모세포제일차감수분렬회복중적작용，용란포자격소（ FSH）、 SB203580（ p38MAPK 억제제）、 PKA 격활제 forskolin 단독작용급 FSH 여SB203580、 FSH여PKA억제제H89공동배양반마어란모세포。채집불동배양시간적란모세포，용SDS-PAGE전영화Western Blot단백질면역인적기술검측p38MAPK린산화，관찰생발포파렬（GVBD）정황。결과표명，반마어란모세포재FSH유발하， p38 MAPK활성명현승고；단독첨가SB203580반마어란모세포p38 MAPK활성수도명현억제，동일시간GVBD발생솔균저우대조조이급동시첨가FSH여SB203580적실험조； FSH여SB203580공동작용조반마어란모세포， GVBD발생솔여SB203580실험조차이성불현저，여FSH조상비현저강저。용H89여FSH공동배양란모세포시，수연p38MAPK잉피격활단량흔소。용forskolin단독배양란모세포， p38MAPK능피격활，차비대조조명현승고。결과제시，재FSH유도적반마어란모세포감수분렬회복과정중p38MAPK피격활，차p38MAPK적격활시PKA의뢰성적。
To clarify the effects of p38 mitogen-activated protein kinase (p38MAPK) on the resumption of the first meio-sis in Danio rerio oocytes, the oocytes were cultured with the following chemicals: no chemical, FSH, SB203580 (p38MAPK inhibitors), forskolin (PKA activator), FSH+SB203580 and FSH+H89 (PKA inhibitor).The oocytes were sampled at different culture times , p38 MAPK phosphorylation was examined with SDS -PAGE electrophoresis and West-ern Blot and the germinal vesicle breakdown ( GVBD) of oocytes was observed.The results showed that the p38MAPK activ-ities increased significantly when the oocytes were induced by FSH.But p38MAPK activities in D.rerio oocytescultured with SB203580 was inhibited obviously and the GVBD rate was lower than control group and the group cultured with FSH and SB203580.The GVBD rates had no significant differences between the group of oocytes cultured with both of FSH and SB203580 and that of oocytes cultured with only SB 203580, but the former group was significantly reduced compared to FSH group.p38 MAPK activity in oocytes in response to the H 89 and FSH induction was still happened , with little a-mount.The p38MAPK can be activated after joining the forskolin without FSH , and it was significantly higher than the con-trol group.Results showed that p38MAPK was activated under the induction of FSH during the resumption of meiosis in D.rerio oocytes.And p38MAPK activation is PKA-dependent.