Niu Yuyu, Yang Shihua, Yu Yang, Ding Chenhui, Yang Jifeng, Wang Shufen, Ji Shaohui, He Xiechao, Xie Yunhua, Tang Xianghui, Zhou Qi, Ji Weizhi
Kunming Primate Research Center & Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan, People's Republic of China.
Cloning Stem Cells. 2008 Mar;10(1):25-36. doi: 10.1089/clo.2007.0040.
Somatic cell nuclear transfer (SCNT) is a remarkable process in which a somatic cell nucleus is acted upon by the ooplasm via mechanisms that today remain unknown. Here we show the developmental competence (% blastocyst) of embryos derived from SCNT (21%) was markedly (p < 0.05) impaired compared with those derived from in vitro fertilization (IVF) (42.1%) in rhesus monkey. Also, SCNT embryos were abnormal in their time course of embryonic development. SCNT produced embryos reached the eight-cell stage faster than did IVF produced embryos. We compare the transcription patterns of five nucleolar-related proteins-nucleolin, nucleophosmin, fibrillarin, PAF53, and UBF-in single IVF and SCNT blastocysts by RT-PCR. The SCNT embryos showed abnormal gene transcription. Immunolocalization of fibrillarin was undetectable in 8-cell and 16-cell SCNT embryos, indicating embryonic genomic activation was delayed in monkey embryos produced by SCNT compared to their IVF-derived counterparts. Some of SCNT embryos appeared to relative higher developmental potential and fibrillarin expression by prolonged exposure of incoming nuclei to a cytoplasm. Thus, our data show that SCNT embryos are characterized by abnormal cleavage and the timely onset of embryonic genome transcription, deficits that may explain their reduced pre- and postimplantation developmental capacity.
体细胞核移植（SCNT）是一个非凡的过程，在这个过程中，体细胞核通过至今仍未知的机制受到卵细胞质的作用。在这里，我们表明，与恒河猴体外受精（IVF）产生的胚胎（42.1%）相比，体细胞核移植产生的胚胎的发育能力（囊胚率21%）明显受损（p < 0.05）。此外，体细胞核移植胚胎在胚胎发育的时间进程上也不正常。体细胞核移植产生的胚胎比体外受精产生的胚胎更快地达到八细胞阶段。我们通过逆转录聚合酶链反应（RT-PCR）比较了单个体外受精和体细胞核移植囊胚中五种核仁相关蛋白——核仁素、核磷蛋白、原纤维蛋白、PAF53和上游结合因子（UBF）的转录模式。体细胞核移植胚胎表现出异常的基因转录。在八细胞和十六细胞的体细胞核移植胚胎中未检测到原纤维蛋白的免疫定位，这表明与体外受精来源的胚胎相比，体细胞核移植产生的猴胚胎的胚胎基因组激活延迟。通过将导入的细胞核长时间暴露于细胞质中，一些体细胞核移植胚胎似乎具有相对较高的发育潜力和原纤维蛋白表达。因此，我们的数据表明，体细胞核移植胚胎的特征是卵裂异常和胚胎基因组转录的及时启动，这些缺陷可能解释了它们在植入前和植入后发育能力的降低。
