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国家重点基础研究发展计划(001CB509903)

作品数:4 被引量:14H指数:2
相关作者:李瑞珍邵华谢颖颖郑俊克马恒东更多>>
相关机构:上海交通大学医学院附属新华医院四川农业大学上海交通大学更多>>
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Nuclear reprogramming: the zygotic transcription program is established through an“erase-and-rebuild” strategy被引量:6
2007年
Oocytes display a maternal-specific gene expression profile, which is switched to a zygotic profile when a haploid set of chromatin is passed on to the fertilized egg that develops into an embryo. The mechanism underlying this transcription reprogramming is currently unknown. Here we demonstrate that by the time when transcription is shut down in germinal vesicle oocytes, a range of general transcription factors and transcriptional regulators are dissociated from the chromatin. The global dissociation of chromatin factors (CFs) disrupts physical contacts between the chromatin and CFs and leads to erasure of the maternal transcription program at the functional level. Critical transcription factors and regulators remain separated from chromatin for a prolonged period, and become re-associated with chromatin shortly after pronuclear formation. This is followed temporally by the re-establishment of nuclear functions such as DNA replication and transcription. We propose that the maternal transcription program is erased during oogenesis to generate a relatively naive chromatin and the zygotic transcription program is rebuilt de novo after fertilization. This process is termed as the "erase-and-rebuild" process, which is used to reset the transcription program, and most likely other nuclear processes as well, from a maternal one to that of the embryo. We further show in the accompanying paper (Gao T, et al., Cell Res 2007; 17:135-150.) that the same strategy is also employed to reprogram transcriptional profiles in somatic cell nuclear transfer and parthenogenesis, suggesting that this model is universally applicable to all forms of transcriptional reprogramming during early embryogenesis. Displacement of CFs from chromatin also offers an explanation for the phenomenon of transcription silence during the maternal to zygotic transition.
Feng SunHaiyan FangRuizhen LiTianlong GaoJunke ZhengXuejin ChenWenqin YingHui Z Sheng
关键词:TRANSCRIPTIONEMBRYOGENESISFERTILIZATIONOOGENESIS
不同的固定及渗透方法对小鼠卵母细胞免疫荧光染色的影响被引量:6
2008年
目的探索有效的染色前固定及渗透卵母细胞的方法,以提高小鼠卵母细胞免疫荧光染色的效率。方法采用4种不同的固定及渗透方法处理小鼠卵母细胞:①方案A:先用4%多聚甲醛(PFA)固定1 h,再用0.5%Triton X-100穿透10 min;②方案B:先用0.5%Triton X-100穿透10 min,再用4%PFA固定1 h;③方案C:用1%PFA+0.5%Triton X-100同时固定与穿透1 h;④方案D:用-20℃预冷甲醇固定15 min。分析10种核蛋白(TFⅡA、TFⅡB、TAF1、TAF4、polⅡ、BRF1、MeCP2、MBD2ab、HP1α及HP1β)在小鼠卵母细胞中的免疫荧光染色效果。结果只有方案C能够完全检测出这10种蛋白质。结论用1%PFA+0.5%Triton X-100同时固定与穿透1 h,可以在小鼠卵母细胞形态保持良好的同时检测出更多的蛋白质。
谢颖颖邵华郑俊克马恒东李瑞珍
关键词:卵母细胞免疫荧光染色激光扫描共聚焦显微镜小鼠
胚胎干细胞向神经细胞的诱导分化及其在神经系统退行性疾病的应用
2007年
胚胎干细胞具有向三个胚层细胞分化的潜能,被视为治疗多种疾病包括神经系统退行性疾病的一种新兴手段。在现阶段,通过不同的诱导途径可将小鼠和人胚胎干细胞诱导成为神经前体细胞和特定类型的神经元,但现有的方法诱导得到的神经细胞比例较低且不易分拣出来。进一步解决这些问题,将为神经系统退行性疾病的临床治疗带来新的希望。
周君梅褚建新陈学进
关键词:胚胎干细胞诱导分化神经系统退行性疾病
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