雌激素受体α抑制剂MPP对小鼠合子型基因组激活的影响可能与S期启动相关

发布时间：2018-01-18 16:01

本文关键词：雌激素受体α抑制剂MPP对小鼠合子型基因组激活的影响可能与S期启动相关　出处：《中国组织化学与细胞化学杂志》2015年06期 　论文类型：期刊论文

【摘要】：目的观察雌激素受体α(ERα)特异性抑制剂甲基哌啶吡唑(Methyl-Piperidino-Pyrazole,MPP)对小鼠早期胚胎发育的影响与合子型基因组激活(ZGA)发生的时间是否相关,并检测与小鼠ZGA密切相关的2-细胞胚DNA复制是否受MPP处理的影响,为进一步探讨ERα在小鼠ZGA过程中的作用机制提供实验基础。方法 1收集昆明小鼠1-细胞胚、2-细胞胚和4-细胞胚,以空白KSOM培养液(单纯型优化的胚胎培养基)为对照组,以KSOM中添加MPP为实验组;将各时间点收集的早胚培养3h后再移入新KSOM培养液中继续培养至囊胚阶段,并记录囊胚数;2收集昆明小鼠末期1-细胞胚,分别置于上述两组培养液中培养15h至2-细胞胚S期后期,用荧光显微镜技术检测2-细胞胚核像变化;Brd U标记法分析DNA复制情况。结果1于不同时间段经MPP 3h处理后,在1-细胞胚末期、2-细胞早期和2-细胞胚中后期,MPP处理对小鼠早胚发育具有显著的抑制作用,其阶段特异性影响与小鼠ZGA发生的时间一致。另外,MPP对2-细胞胚G1/S过渡期(36~39h)抑制明显,但是对S期中后期(39~42h)进程无显著影响。2与KSOM组相比,经MPP处理15h后,2-细胞胚核变圆;Brd U标记水平明显下降。结论MPP在小鼠早胚中的阶段性作用与ZGA有关;ERα可能通过促进S期启动来调控ZGA。
[Abstract]:Objective to observe methylpiperidopyrazole methyl-Piperidino-Pyrazole, a specific inhibitor of estrogen receptor 伪 (ER 伪). The effect of MPP on the development of mouse early embryos was related to the time of ZGA (zygotic genome activation). The effects of MPP treatment on DNA replication of 2-cell embryos closely related to mouse ZGA were also detected. In order to further explore the mechanism of ER 伪 in mouse ZGA. Methods 1 Kunming mouse 1-cell embryos, 2-cell embryos and 4- cell embryos were collected. Blank KSOM medium (simple optimized embryo medium) was used as control group, and MPP was added to KSOM as experimental group. The early embryos collected at each time point were cultured for 3 hours and then transferred into the new KSOM culture medium to continue to culture to the blastocyst stage, and the number of blastocysts was recorded. (2) the late 1-cell embryos of Kunming mice were collected and cultured in the above two groups for 15 hours to the late S phase of 2-cell embryos, and the nuclear images of 2-cell embryos were detected by fluorescence microscopy. Results 1 after being treated with MPP for 3 h at different time periods, DNA replication was analyzed by Brd U labeling method. Results 1 after 3 hours of MPP treatment, the early stage of 1-cell embryo and the middle and late stage of 2-cell embryo were observed at the end of 1-cell embryo. MPP treatment had a significant inhibitory effect on early embryo development in mice, and its phase specific effect was consistent with the time of occurrence of ZGA in mice. MPP inhibited the G1 / S transition period of 2-cell embryos for 36 ~ 39 h), but had no significant effect on the process of 39 ~ 39 ~ 42 h) in the middle and late S phase. 2 compared with the KSOM group, there was no significant difference between the two groups. After 15 hours of MPP treatment, the nuclei of 2-cell embryos became round. Conclusion the stage effect of MPP in mouse early embryo is related to ZGA. ER 伪 may regulate ZGA by promoting S phase priming.
【作者单位】：福建医科大学基础医学院人体解剖学与组织胚胎学系;福建医科大学基础医学院细胞与发育工程研究中心;
【基金】：国家自然科学基金资助项目(81170624)
【分类号】：R321
【正文快照】： 小鼠合子型基因组激活(zygotic gene activation,ZGA)发生于1-细胞末期至2-细胞早期和2-细胞中后期[1],在此期间,合子型基因产物逐渐取代母源因子,为早胚的后续发育奠定基础(图1)[2]。因此,ZGA是继受精之后的一个重要的分子事件。然而ZGA发生的具体机制尚未清楚。本课题组前期

【相似文献】