OCT-4 expression is essential for the segregation of trophectoderm lineages in porcine preimplantation embryos.
OCT-4 expression is essential for the segregation of trophectoderm lineages in porcine preimplantation embryos.
复制标题
DOI：
10.1262/jrd.2016-040
复制
发表时间：
2016-08-25
期刊：
影响因子：
--
通讯作者：
Sawai K
中科院分区：
文献类型：
--
作者：
Emura N;Sakurai N;Takahashi K;Hashizume T;Sawai K
Oct-4是POU转录因子家族中的一员，是调节小鼠内细胞团(ICM)和滋养外胚层(TE)从桑椹胚向囊胚转变过程中分离的关键因子。然而，对其在猪早期胚胎发育中的作用却知之甚少。为了确定Oct-4在猪胚胎ICM和TE分离中的作用，我们利用RNA干扰技术对猪胚胎的发育形态进行了研究。我们的实验表明，当1-细胞期胚胎与靶向敲除Oct-4(Oct-4-siRNA)的小干扰RNA(SiRNA)和四甲基罗丹明-异硫氰酸酯(TRITC)-葡聚糖偶联物(Dx)共同注射时，它们不能形成囊胚。因此，在本研究中，我们构建了包含卵裂球的嵌合胚胎，这些卵裂球要么正常表达Oct-4，要么通过将Oct-4-siRNA和Dx共同注射到2-4细胞期胚胎的一个卵裂球中下调Oct-4的表达。在同时注射对照siRNA和Dx的对照胚胎中，Dx阳性细胞在几乎所有被检查的囊胚中都参与了TE谱系。相比之下，来自OCT-4-siRNA和Dx联合注射的卵裂球的Dx阳性细胞在几乎一半的囊胚中退化。这可能是由于这些细胞不能分化为TE谱系所致。实时定量RT-PCR分析表明，在注射Oct-4-siRNA的桑椹胚中，SOX2、TEAD4、FGF4和FGFR1-IIIc的表达水平与对照组没有差异，这些都是由Oct-4调控的。然而，CDX2是一种在TE谱系中特异表达的分子，前者的水平显著高于后者。我们的结果表明，Oct-4在卵裂球中的持续表达是猪胚胎形成所必需的。
Oct-4, a member of the POU family of transcription factors, is a key factor that regulates the segregation of the inner cell mass (ICM) and the trophectoderm (TE) during the transition from morula to blastocyst in mice. However, little is known about its role in porcine early embryogenesis. To determine the function of OCT-4 in the ICM and TE segregation of porcine embryos, we studied the developmental morphology of porcine embryos using RNA interference technology. Our experiments demonstrated that when 1-cell stage embryos were co-injected with the small interfering RNA (siRNA)for targeted knockdown of OCT-4 (OCT-4-siRNA) and tetramethylrhodamine isothiocyanate (TRITC)-dextran conjugate (Dx), they failed to form blastocysts. Therefore, in this study, we constructed chimeric embryos comprising blastomeres that either expressed OCT-4 normally or showed downregulated OCT-4 expression by co-injection of OCT-4-siRNA and Dx into one blastomere in 2- to 4-cell stage embryos. In control embryos, which were co-injected with control siRNA and Dx, Dx-positive cells contributed to the TE lineage in almost all the blastocysts examined. In contrast, Dx-positive cells derived from a blastomere co-injected with OCT-4-siRNA and Dx were degenerated in almost half the blastocysts. This was probably due to the inability of these cells to differentiate into the TE lineage. Real-time RT-PCR analysis revealed no difference in the levels of SOX2, TEAD4, FGF4 and FGFR1-IIIc, all of which are known to be regulated by OCT-4, between the OCT-4-siRNA-injected morulae and the control ones. However, the level of CDX2, a molecule specifically expressed in the TE lineage, was significantly higher in the former than in the latter. Our results indicate that continuous expression of OCT-4 in blastomeres is essential for TE formation of porcine embryos.
登录
查看更多内容
影响因子：
4.4
作者：
Cao S;Han J;Wu J;Li Q;Liu S;Zhang W;Pei Y;Ruan X;Liu Z;Wang X;Lim B;Li N
通讯作者：
Li N
DOI：
10.1016/j.bbrc.2006.08.004
发表时间：
2006-10-06
影响因子：
3.1
作者：
Lee, Eugine;Lee, So Hyun;Lee, Byeong Chun
通讯作者：
Lee, Byeong Chun
影响因子：
64.5
作者：
Niwa, H;Toyooka, T;Rossant, J
通讯作者：
Rossant, J
影响因子：
2.8
作者：
du Puy, Leonie;Lopes, Susana M. Chuva de Sousa;Roelen, Bernard A. J.
通讯作者：
Roelen, Bernard A. J.
影响因子：
3.8
作者：
Kunath, T;Strumpf, D;Rossant, J
通讯作者：
Rossant, J