Analysis of nuclear reprogramming in cloned miniature pig embryos by expression of Oct-4 and Oct-4 related genes
Analysis of nuclear reprogramming in cloned miniature pig embryos by expression of Oct-4 and Oct-4 related genes
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DOI：
10.1016/j.bbrc.2006.08.004
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发表时间：
2006-10-06
影响因子：
3.1
通讯作者：
Lee, Byeong Chun
中科院分区：
文献类型：
--
作者：
Lee, Eugine;Lee, So Hyun;Lee, Byeong Chun
异种移植是一个迅速发展的研究领域，克隆小型猪被认为是异种移植的模式动物。然而，体细胞核移植(SCNT)的效率极低，大多数克隆导致早期死亡和几种异常发育。SCNT胚胎发育失败的一个可能解释是卵母细胞对体细胞核的重新编程不足。为了验证这一点，我们用与胚胎发育有关的OCT-4和OCT-4相关基因(Ndp52II、Dppa2、Dppa3和Dppa5)、胎盘发育重要的Hand1和GATA-4作为分子标记，利用RT-PCR技术分析了分化的成纤维细胞核和胚胎生殖细胞核的重编程能力。在来自成纤维细胞的克隆孵化囊胚中，Oct-4的表达水平显著降低(P<；0.05)，许多来自成纤维细胞的克隆未能重新激活至少一个测试基因，而大多数生殖细胞克隆和对照胚胎正确地表达了这些基因。综上所述，我们的结果表明，成纤维细胞来源的克隆胚胎的重编程是高度异常的，这种不适当的重编程可能是体细胞来源克隆早期致死和植入后异常的原因之一。(C)2006 Elsevier Inc.保留所有权利。
Xenotransplantation is a rapidly expanding field of research and cloned miniature pigs have been considered as a model animal for it. However, the efficiency of somatic cell nuclear transfer (SCNT) is extremely low, with most clones resulting in early lethality and several kinds of aberrant development. A possible explanation for the developmental failure of SCNT embryos is insufficient reprogramming of the somatic cell nucleus by the oocyte. In order to test this, we analyzed the reprogramming capacity of differentiated fibroblast cell nuclei and embryonic germ cell nuclei with Oct-4 and Oct-4 related genes (Ndp52II, Dppa2, Dppa3, and Dppa5), which are important for embryonic development, Hand1 and GATA-4, which are important for placental development, as molecular markers using RT-PCR. The Oct-4 expression level was significantly lower (P < 0.05) in cloned hatched blastocysts derived from fibroblasts and many of fibroblast-derived clones failed to reactivate at least one of the tested genes, while most of the germ cell clones and control embryos correctly expressed these genes. In conclusion, our results suggest that the reprogramming of fibroblast-derived cloned embryos is highly aberrant and this improper reprogramming could be one reason of the early lethality and post-implantation anomalies of somatic cell-derived clones. (c) 2006 Elsevier Inc. All rights reserved.