Nanog co-regulated by Nodal/Smad2 and Oct4 is required for pluripotency in developing mouse epiblast

• DOI: 10.1016/j.ydbio.2014.06.002
• Publication Date: 2014-06-12T01:15:50Z
• Authors: Liang Tso Sun; Shinpei Yamaguchi; Kunio Hirano; Tomoko Ichisaka; Takao Kuroda; Takashi Tada
• Keywords: Pluripotency; Pluripotent Stem Cells; 0301 basic medicine; Chromatin Immunoprecipitation; Nodal Protein; Nodal; Oct4; Smad2 Protein; Nanog; Models, Biological; Cell Line; Mice; 03 medical and health sciences; Animals; Luciferases; Molecular Biology; Homeodomain Proteins; Epiblast; Reverse Transcriptase Polymerase Chain Reaction; Cell Biology; Nanog Homeobox Protein; Immunohistochemistry; Gene Knockdown Techniques; Octamer Transcription Factor-3; Smad2; Germ Layers; Developmental Biology

Nanog, a core pluripotency factor, is required for stabilizing pluripotency of inner cell mass (ICM) and embryonic stem cells (ESCs), and survival of primordial germ cells in mice. Here, we have addressed function and regulation of Nanog in epiblasts of postimplantation mouse embryos by conditional knockdown (KD), chromatin immunoprecipitation (ChIP) using in vivo epiblasts, and protein interaction with the Nanog promoter in vitro. Differentiation of Nanog-KD epiblasts demonstrated requirement for Nanog in stabilization of pluripotency. Nanog expression in epiblast is directly regulated by Nodal/Smad2 pathway in a visceral endoderm-dependent manner. Notably, Nanog promoters switch from Oct4/Esrrb in ICM/ESCs to Oct4/Smad2 in epiblasts. Smad2 directly associates with Oct4 to form Nanog promoting protein complex. Collectively, these data demonstrate that Nanog plays a key role in stabilizing Epiblast pluripotency mediated by Nodal/Smad2 signaling, which is involved in Nanog promoter switching in early developing embryos.