title: Nanog co-regulated by Nodal/Smad2 and Oct4 is required for pluripotency in developing mouse epiblast.
abstract: 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.
authors.affiliation: Department of Stem Cell Engineering, Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Liang Tso
authors.initials: LT
authors.lastname: Sun
authors.affiliation: Department of Stem Cell Engineering, Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Shinpei
authors.initials: S
authors.lastname: Yamaguchi
authors.affiliation: Department of Stem Cell Engineering, Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Kunio
authors.initials: K
authors.lastname: Hirano
authors.affiliation: Department of Reprogramming Science, Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Ichisaka
authors.affiliation: Laboratory of Neural Differentiation, Graduate School of Brain Science, Doshisha University, Kyoto 619-0225, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Kuroda
authors.affiliation: Department of Stem Cell Engineering, Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, Japan. Electronic address: ttada@frontier.kyoto-u.ac.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Tada
mesh_terms: D000818:Animals; D002460:Cell Line; D047369:Chromatin Immunoprecipitation; D055785:Gene Knockdown Techniques; D005855:Germ Layers / Q000196:embryology* / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D007150:Immunohistochemistry; D008156:Luciferases; D051379:Mice; D008954:Models, Biological; D000071317:Nanog Homeobox Protein; D055457:Nodal Protein / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000378:metabolism / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051899:Smad2 Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D018398:Homeodomain Proteins; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D055457:Nodal Protein; C526008:Nodal protein, mouse; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D051899:Smad2 Protein; C497783:Smad2 protein, mouse; D008156:Luciferases
keywords: Epiblast; Nanog; Nodal; Oct4; Pluripotency; Smad2