本文關(guān)鍵詞：雌激素受體α抑制劑MPP對(duì)小鼠合子型基因組激活的影響可能與S期啟動(dòng)相關(guān)　出處：《中國(guó)組織化學(xué)與細(xì)胞化學(xué)雜志》2015年06期 　論文類型：期刊論文

  更多相關(guān)文章： 雌激素受體α 抑制劑 小鼠 合子型基因組激活 細(xì)胞周期

【摘要】：目的觀察雌激素受體α(ERα)特異性抑制劑甲基哌啶吡唑(Methyl-Piperidino-Pyrazole,MPP)對(duì)小鼠早期胚胎發(fā)育的影響與合子型基因組激活(ZGA)發(fā)生的時(shí)間是否相關(guān),并檢測(cè)與小鼠ZGA密切相關(guān)的2-細(xì)胞胚DNA復(fù)制是否受MPP處理的影響,為進(jìn)一步探討ERα在小鼠ZGA過(guò)程中的作用機(jī)制提供實(shí)驗(yàn)基礎(chǔ)。方法 1收集昆明小鼠1-細(xì)胞胚、2-細(xì)胞胚和4-細(xì)胞胚,以空白KSOM培養(yǎng)液(單純型優(yōu)化的胚胎培養(yǎng)基)為對(duì)照組,以KSOM中添加MPP為實(shí)驗(yàn)組;將各時(shí)間點(diǎn)收集的早胚培養(yǎng)3h后再移入新KSOM培養(yǎng)液中繼續(xù)培養(yǎng)至囊胚階段,并記錄囊胚數(shù);2收集昆明小鼠末期1-細(xì)胞胚,分別置于上述兩組培養(yǎng)液中培養(yǎng)15h至2-細(xì)胞胚S期后期,用熒光顯微鏡技術(shù)檢測(cè)2-細(xì)胞胚核像變化;Brd U標(biāo)記法分析DNA復(fù)制情況。結(jié)果1于不同時(shí)間段經(jīng)MPP 3h處理后,在1-細(xì)胞胚末期、2-細(xì)胞早期和2-細(xì)胞胚中后期,MPP處理對(duì)小鼠早胚發(fā)育具有顯著的抑制作用,其階段特異性影響與小鼠ZGA發(fā)生的時(shí)間一致。另外,MPP對(duì)2-細(xì)胞胚G1/S過(guò)渡期(36~39h)抑制明顯,但是對(duì)S期中后期(39~42h)進(jìn)程無(wú)顯著影響。2與KSOM組相比,經(jīng)MPP處理15h后,2-細(xì)胞胚核變圓;Brd U標(biāo)記水平明顯下降。結(jié)論MPP在小鼠早胚中的階段性作用與ZGA有關(guān);ERα可能通過(guò)促進(jìn)S期啟動(dòng)來(lái)調(diào)控ZGA。
[Abstract]:Objective to observe methylpiperidopyrazole methyl-Piperidino-Pyrazole, a specific inhibitor of estrogen receptor 偽 (ER 偽). 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 偽 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 偽 may regulate ZGA by promoting S phase priming.
【作者單位】： 福建醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院人體解剖學(xué)與組織胚胎學(xué)系;福建醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院細(xì)胞與發(fā)育工程研究中心;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(81170624)
【分類號(hào)】：R321
【正文快照】： 小鼠合子型基因組激活(zygotic gene activation,ZGA)發(fā)生于1-細(xì)胞末期至2-細(xì)胞早期和2-細(xì)胞中后期[1],在此期間,合子型基因產(chǎn)物逐漸取代母源因子,為早胚的后續(xù)發(fā)育奠定基礎(chǔ)(圖1)[2]。因此,ZGA是繼受精之后的一個(gè)重要的分子事件。然而ZGA發(fā)生的具體機(jī)制尚未清楚。本課題組前期

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