Homeodomain transcription factor Nanog plays a critical role in maintaining murine embryonic stem (ES) cells pluripotency. However, its expression regulation largely remains unknown. In this study we show that Nogo receptor (NgR) participates in the regulation of Nanog expression via Stat3 pathway. Activation of NgR results in the phosphorylation of Stat3 and increases expression levels of Nanog mRNA and protein, which inhibits differentiation of embryoid bodies. This up-regulation of Nanog can be abolished by NgR inhibitor PI-PLC and NEP1-40, or phospho-Stat3 inhibitor AG490 and rapamycin. Immunofluorescence assay demonstrates that NgR and its ligand Nogo-A/B exist on mouse blastocysts and cultured ES cells, suggesting NgR might play a role in early embryo development.
Get full access to this article
View all access options for this article.
References
1.
EvansMJ and KaufmanMH. (1981). Establishment in culture of pluripotential cells from mouse embryos. Nature, 292:154–156.
2.
MartinGR. (1981). Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc Natl Acad Sci USA, 78:7634–7638.
3.
MitsuiK, TokuzawaY, ItohH, SegawaK, MurakamiM, TakahashiK, MaruyamaM, MaedaM and YamanakaS. (2003). The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell, 113:631–642.
4.
ChambersI, ColbyD, RobertsonM, NicholsJ, LeeS, TweedieS and SmithA. (2003). Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell, 113:643–655.
5.
KurodaT, TadaM, KubotaH, KimuraH, HatanoSY, SuemoriH, NakatsujiN and TadaT. (2005). Octamer and Sox elements are required for transcriptional cis regulation of Nanog gene expression. Mol Cell Biol, 25:2475–2485.
6.
RoddaDJ, ChewJL, LimLH, LohYH, WangB, NgHH and RobsonP. (2005). Transcriptional regulation of nanog by OCT4 and SOX2. J Biol Chem, 280:24731–24737.
7.
RichardsM, TanSP, TanJH, ChanWK and BongsoA. (2004). The transcriptome profile of human embryonic stem cells as defined by SAGE. Stem Cells (Dayton, Ohio), 22:51–64.
BurdonT, SmithA and SavatierP. (2002). Signalling, cell cycle and pluripotency in embryonic stem cells. Trends Cell Biol, 12:432–438.
10.
BabaieY, HerwigR, GreberB, BrinkTC, WruckW, GrothD, LehrachH, BurdonT and AdjayeJ. (2007). Analysis of Oct4-dependent transcriptional networks regulating self-renewal and pluripotency in human embryonic stem cells. Stem Cells (Dayton, Ohio), 25:500–510.
11.
LinT, ChaoC, SaitoS, MazurSJ, MurphyME, AppellaE and XuY. (2005). p53 induces differentiation of mouse embryonic stem cells by suppressing Nanog expression. Nat Cell Biol, 7:165–171.
12.
PereiraL, YiF and MerrillBJ. (2006). Repression of Nanog gene transcription by Tcf3 limits embryonic stem cell self-renewal. Mol Cell Biol, 26:7479–7491.
13.
SatoN, SanjuanIM, HekeM, UchidaM, NaefF and BrivanlouAH. (2003). Molecular signature of human embryonic stem cells and its comparison with the mouse. Dev Biol, 260:404–413.
14.
SatoN, MeijerL, SkaltsounisL, GreengardP and BrivanlouAH. (2004). Maintenance of pluripotency in human and mouse embryonic stem cells through activation of Wnt signaling by a pharmacological GSK-3-specific inhibitor. Nat Med, 10:55–63.
15.
StormMP, BoneHK, BeckCG, BourillotPY, SchreiberV, DamianoT, NelsonA, SavatierP and WelhamMJ. (2007). Regulation of Nanog expression by phosphoinositide 3-kinase-dependent signaling in murine embryonic stem cells. J Biol Chem, 282:6265–6273.
16.
NiwaH, BurdonT, ChambersI and SmithA. (1998). Self-renewal of pluripotent embryonic stem cells is mediated via activation of STAT3. Genes Dev, 12:2048–2060.
17.
RazR, LeeCK, CannizzaroLA, d’EustachioP, and LevyDE (1999). Essential role of STAT3 for embryonic stem cell pluripotency. Proc Natl Acad Sci USA, 96:2846–2851.
18.
PanG and ThomsonJA. (2007). Nanog and transcriptional networks in embryonic stem cell pluripotency. Cell Res, 17:42–49.
19.
SuzukiA, RayaA, KawakamiY, MoritaM, MatsuiT, NakashimaK, GageFH, Rodriguez-EstebanC and Izpisua BelmonteJC. (2006). Nanog binds to Smad1 and blocks bone morphogenetic protein-induced differentiation of embryonic stem cells. Proc Natl Acad Sci USA, 103:10294–10299.
20.
ChenMS, HuberAB, van der HaarME, FrankM, SchnellL, SpillmannAA, ChristF and SchwabME. (2000). Nogo-A is a myelin-associated neurite outgrowth inhibitor and an antigen for monoclonal antibody IN-1. Nature, 403:434–439.
21.
GrandPreT, NakamuraF, VartanianT and StrittmatterSM. (2000). Identification of the Nogo inhibitor of axon regeneration as a reticulon protein. Nature, 403:439–444.
22.
FournierAE, GrandPreT and StrittmatterSM. (2001). Identification of a receptor mediating Nogo-66 inhibition of axonal regeneration. Nature, 409:341–346.
23.
HuberAB, WeinmannO, BrosamleC, OertleT and SchwabME. (2002). Patterns of Nogo mRNA and protein expression in the developing and adult rat and after CNS lesions. J Neurosci, 22:3553–3567.
24.
AcevedoL, YuJ, Erdjument-BromageH, MiaoRQ, KimJE, FultonD, TempstP, StrittmatterSM and SessaWC. (2004). A new role for Nogo as a regulator of vascular remodeling. Nat Med, 10:382–388.
25.
LiQ, QiB, OkaK, ShimakageM, YoshiokaN, InoueH, HakuraA, KodamaK, StanbridgeEJ and YutsudoM. (2001). Link of a new type of apoptosis-inducing gene ASY/Nogo-B to human cancer. Oncogene, 20:3929–3936.
26.
QiB, QiY, WatariA, YoshiokaN, InoueH, MinemotoY, YamashitaK, SasagawaT and YutsudoM. (2003). Pro-apoptotic ASY/Nogo-B protein associates with ASYIP. J Cell Physiol, 196:312–318.
27.
OertleT, MerklerD and SchwabME. (2003). Do cancer cells die because of Nogo-B?. Oncogene, 22:1390–1399.
28.
YiuG and HeZ. (2003). Signaling mechanisms of the myelin inhibitors of axon regeneration. Curr Opin Neurobiol, 13:545–551.
29.
FilbinMT. (2003). Myelin-associated inhibitors of axonal regeneration in the adult mammalian CNS. Nat Rev, 4:703–713.
30.
WangB, XiaoZ, ChenB, HanJ, GaoY, ZhangJ, ZhaoW, WangX and DaiJ. (2008). Nogo-66 promotes the differentiation of neural progenitors into astroglial lineage cells through mTOR-STAT3 pathway. PLoS ONE, 3:e1856.
31.
StewartR, StojkovicM and LakoM. (2006). Mechanisms of self-renewal in human embryonic stem cells. Eur J Cancer, 42:1257–1272.
32.
RaoM. (2004). Conserved and divergent paths that regulate self-renewal in mouse and human embryonic stem cells. Dev Biol, 275:269–286.
33.
HyslopL, StojkovicM, ArmstrongL, WalterT, StojkovicP, PrzyborskiS, HerbertM, MurdochA, StrachanT and LakoM. (2005). Downregulation of NANOG induces differentiation of human embryonic stem cells to extraembryonic lineages. Stem Cells (Dayton, Ohio), 23:1035–1043.
34.
HamazakiT, OkaM, YamanakaS and TeradaN. (2004). Aggregation of embryonic stem cells induces Nanog repression and primitive endoderm differentiation. J Cell Sci, 117:5681–5686.
35.
HaoJ, LiTG, QiX, ZhaoDF and ZhaoGQ. (2006). WNT/beta-catenin pathway up-regulates Stat3 and converges on LIF to prevent differentiation of mouse embryonic stem cells. Dev Biol, 290:81–91.
36.
YingQL, NicholsJ, ChambersI and SmithA. (2003). BMP induction of Id proteins suppresses differentiation and sustains embryonic stem cell self-renewal in collaboration with STAT3. Cell, 115:281–292.
