Gao Tianlong, Zheng Junke, Xing Fengying, Fang Haiyan, Sun Feng, Yan Ayong, Gong Xun, Ding Hui, Tang Fan, Sheng Hui Z
Program for Graduation Studies, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Cell Res. 2007 Feb;17(2):135-50. doi: 10.1038/cr.2007.2.
Somatic cell nuclear transfer (SCNT) and parthenogenesis are alternative forms of reproduction and development, building new life cycles on differentiated somatic cell nuclei and duplicated maternal chromatin, respectively. In the preceding paper (Sun F, et al., Cell Res 2007; 17:117-134.), we showed that an "erase-and-rebuild" strategy is used in normal development to transform the maternal gene expression profile to a zygotic one. Here, we investigate if the same strategy also applies to SCNT and parthenogenesis. The relationship between chromatin and chromatin factors (CFs) during SCNT and parthenogenesis was examined using immunochemical and GFP-fusion protein assays. Results from these studies indicated that soon after nuclear transfer, a majority of CFs dissociated from somatic nuclei and were redistributed to the cytoplasm of the egg. The erasure process in oogenesis is recaptured during the initial phase in SCNT. Most CFs entered pseudo-pronuclei shortly after their formation. In parthenogenesis, all parthenogenotes underwent normal oogenesis, and thus had removed most CFs from chromosomes before the initiation of development. The CFs were subsequently re-associated with female pronuclei in time and sequence similar to that in fertilized embryos. Based on these data, we conclude that the "erase-and-rebuild" process observed in normal development also occurs in SCNT and in parthenogenesis, albeit in altered fashions. The process is responsible for transcription reprogramming in these procedures. The "erase" process in SCNT is compressed and the efficiency is compromised, which likely contribute to the developmental defects often observed in nuclear transfer (nt) embryos. Furthermore, results from this study indicated that the cytoplasm of an egg contains most, if not all, essential components for assembling the zygotic program and can assemble them onto appropriate diploid chromatin of distinct origins.
体细胞克隆(SCNT)和孤雌生殖是生殖与发育的替代形式,分别基于分化的体细胞核和复制的母本染色质构建新的生命周期。在前一篇论文(Sun F等人,《细胞研究》2007年;17:117 - 134)中,我们表明正常发育过程中采用了一种“擦除并重建”策略,将母本基因表达谱转变为合子基因表达谱。在此,我们研究该策略是否也适用于体细胞克隆和孤雌生殖。利用免疫化学和绿色荧光蛋白融合蛋白检测方法,研究了体细胞克隆和孤雌生殖过程中染色质与染色质因子(CFs)之间的关系。这些研究结果表明,核移植后不久,大多数CFs从体细胞核解离并重新分布到卵细胞质中。体细胞克隆初始阶段重现了卵子发生过程中的擦除过程。大多数CFs在伪原核形成后不久进入其中。在孤雌生殖中,所有孤雌胚胎都经历了正常的卵子发生,因此在发育开始前已从染色体上移除了大多数CFs。随后,CFs以与受精胚胎相似的时间和顺序重新与雌原核结合。基于这些数据,我们得出结论,正常发育中观察到的“擦除并重建”过程在体细胞克隆和孤雌生殖中也会发生,尽管方式有所改变。该过程负责这些过程中的转录重编程。体细胞克隆中的“擦除”过程被压缩且效率受损,这可能是核移植(nt)胚胎中经常观察到发育缺陷的原因。此外,本研究结果表明,卵细胞质包含组装合子程序的大部分(如果不是全部)必需成分,并且可以将它们组装到不同来源的合适二倍体染色质上。