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First published online November 11, 2009
doi: 10.1242/10.1242/dev.041160

Development 136, 4011-4020 (2009)
Published by The Company of Biologists 2009
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Conditional knockdown of Nanog induces apoptotic cell death in mouse migrating primordial germ cells

Shinpei Yamaguchi1, Kazuki Kurimoto2, Yukihiro Yabuta2, Hiroyuki Sasaki3, Norio Nakatsuji4,5, Mitinori Saitou2,* and Takashi Tada1,6,{dagger},{ddagger}

1 Stem Cell Engineering, Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan
2 Laboratory for Mammalian Germ Cell Biology, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan
3 Department of Integrated Genetics, National Institute of Genetics, Research Organization of Information and Systems, 1111 Yata, Mishima-shi, Shizuoka 411-8540, Japan
4 Development and Differentiation, Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan
5 Institute for Integrated Cell-Material Sciences (iCeMS), Kyoto University, Kyoto 606-8501, Japan
6 JST, CREST, 4-1-8 Hon-cho, Kawaguchi, Saitama 332-0012, Japan

{ddagger} Author for correspondence (ttada{at}frontier.kyoto-u.ac.jp)

Accepted September 29, 2009

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.

Key words: Nanog, Knockdown, Primordial germ cell, Apoptosis, Mouse


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