Conditional knockdown of Nanog induces apoptotic cell death in mouse migrating primordial germ cells
Conditional knockdown of Nanog induces apoptotic cell death in mouse migrating primordial germ cells
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DOI：
10.1242/dev.041160
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发表时间：
2009-12-01
期刊：
影响因子：
4.6
通讯作者：
Tada, Takashi
中科院分区：
文献类型：
--
作者：
Yamaguchi, Shinpei;Kurimoto, Kazuki;Tada, Takashi
多能因子Nanog在胚胎着床期和原始生殖细胞（PGCs）中表达。缺乏纳米颗粒的小鼠胚胎在植入后很快死亡。为了探索Nanog在生殖细胞中的功能，我们在体内用shRNA有条件地敲低Nanog RNA。纳米shRNA转基因（NRi-Tg）小鼠是通过与NRi-Tg胚胎干细胞形成种系嵌合体而产生的。在E12.5 cre诱导的ER-Cre/NRi-Tg和TNAP-Cre/NRi-Tg双转基因胚胎中，碱性磷酸酶阳性和ssea1阳性的PGCs数量显著减少。在E9.5和E10.5迁移的nanog -敲低PGCs中，尽管Oct4表达，但tunel阳性的凋亡细胞死亡在体内和体外都变得突出。对E10.5 nanog敲低的PGCs进行单细胞微阵列分析，发现大量基因显著上调和下调，包括Tial1、Id1和Suz12。这些数据表明，Nanog作为pgc特异性分子网络的保障，在迁移PGCs的增殖和存活中起着关键作用。
The pluripotency factor Nanog is expressed in peri-implantation embryos and primordial germ cells (PGCs). Nanog-deficient mouse embryos die soon after implantation. To explore the function of Nanog in germ cells, Nanog RNA was conditionally knocked down in vivo by shRNA. Nanog shRNA transgenic (NRi-Tg) mice were generated through the formation of germline chimeras with NRi-Tg embryonic stem cells. In E12.5 Cre-induced ER-Cre/NRi-Tg and TNAP-Cre/NRi-Tg double-transgenic embryos, the number of alkaline phosphatase-positive and SSEA1-positive PGCs decreased significantly. In the E9.5 and E10.5 migrating Nanog-knockdown PGCs, TUNEL-positive apoptotic cell death became prominent in vivo and in vitro, despite Oct4 expression. Single-cell microarray analysis of E10.5 Nanog-knockdown PGCs revealed significant up- and downregulation of a substantial number of genes, including Tial1, Id1 and Suz12. These data suggest that Nanog plays a key role in the proliferation and survival of migrating PGCs as a safeguard of the PGC-specific molecular network.