Restricted accessResearch articleFirst published online 2016-10
The Necessity of OCT-4 and CDX2 for Early Development and Gene Expression Involved in Differentiation of Inner Cell Mass and Trophectoderm Lineages in Bovine Embryos
The functions of POU class 5 transcription factor 1 (Oct-4) and caudal-type homeobox 2 (Cdx2) in the differentiation of the murine inner cell mass (ICM) and trophectoderm (TE) have been described in detail. However, little is known about the roles of OCT-4 and CDX2 in preimplantation bovine embryos. To elucidate their functions during early development in bovine embryos, we performed OCT-4 and CDX2 downregulation using RNA interference. We injected OCT-4- or CDX2-specific short interfering RNAs (siRNAs) into bovine zygotes. The rate of blastocyst development of OCT-4-downregulated embryos was lower compared with uninjected or control siRNA-injected embryos. Gene expression analysis revealed decreased CDX2 and fibroblast growth factor 4 expression in OCT-4-downregulated embryos. CDX2-downregulated embryos developed to the blastocyst stage; however, in most cases, blastocoel formation was delayed. Gene expression analysis revealed decreased GATA3 expression and elevated NANOG expression in CDX2-downregulated embryos. In conclusion, OCT-4 and CDX2 are essential for early development and gene expression involved in differentiation of ICM and TE lineages in bovine embryos.
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References
1.
AllenB.L., FillaM.S., and RapraegerA.C. (2001). Role of heparan sulfate as a tissue-specific regulator of FGF-4 and FGF receptor recognition. J. Cell Biol., 155, 845–858.
2.
AmbrosettiD.C., BasilicoC., and DaileyL. (1997). Synergistic activation of the fibroblast growth factor 4 enhancer by Sox2 and Oct-3 depends on protein-protein interactions facilitated by a specific spatial arrangement of factor binding sites. Mol. Cell. Biol., 17, 6321–6329.
3.
BavisterB.D., LeibfriedM.L., and LiebermanG. (1983). Development of preimplantation embryos of the golden hamster in a defined culture medium. Biol. Reprod., 28, 235–247.
4.
BergD.K., SmithC.S., PeartonD.J., WellsD.N., BroadhurstR., DonnisonM., and PfefferP.L. (2011). Trophectoderm lineage determination in cattle. Dev. Cell, 20, 244–255.
5.
BessonnardS., De MotL., GonzeD., BarriolM., DennisC., GoldbeterA., DupontG., and ChazaudC. (2014). Gata6, Nanog and Erk signaling control cell fate in the inner cell mass through a tristable regulatory network. Development, 141, 3637–3648.
6.
ChenL., YabuuchiA., EminliS., TakeuchiA., LuC.W., HochedlingerK., and DaleyG.Q. (2009). Cross-regulation of the Nanog and Cdx2 promoters. Cell Res. 19, 1052–1061.
7.
DegrelleS.A., CampionE., CabauC., PiumiF., ReinaudP., RichardC., RenardJ.P., and HueI. (2005). Molecular evidence for a critical period in mural trophoblast development in bovine blastocysts. Dev. Biol., 288, 448–460.
8.
DietrichJ.E., and HiiragiT. (2007). Stochastic patterning in the mouse pre-implantation embryo. Development, 134, 4219–4231.
9.
FrumT., HalbisenM.A., WangC., AmiriH., RobsonP., and RalstonA. (2013). Oct4 cell-autonomously promotes primitive endoderm development in the mouse blastocyst. Dev. Cell, 25, 610–622.
10.
FujiiT., MoriyasuS., HirayamaH., HashizumeT., and SawaiK. (2010). Aberrant expression patterns of genes involved in segregation of inner cell mass and trophectoderm lineages in bovine embryos derived from somatic cell nuclear transfer. Cell. Reprogram., 12, 617–625.
11.
FujiiT., SakuraiN., OsakiT., IwagamiG., HirayamaH., MinamihashiA., HashizumeT., and SawaiK. (2013). Changes in the expression patterns of the genes involved in the segregation and function of inner cell mass and trophectoderm lineages during porcine preimplantation development. J. Reprod. Dev., 59, 151–158.
12.
GoissisM.D., and CibelliJ.B. (2014). Functional characterization of CDX2 during bovine preimplantation development in vitro. Mol. Reprod. Dev., 81, 962–970.
13.
HallV.J., ChristensenJ., GaoY., SchmidtM.H., and HyttelP. (2009). Porcine pluripotency cell signaling develops from the inner cell mass to the epiblast during early development. Dev. Dyn., 238, 2014–2024.
14.
HomeP., RayS., DuttaD., BronshteynI., LarsonM., and PaulS. (2009). GATA3 is selectively expressed in the trophectoderm of peri-implantation embryo and directly regulates Cdx2 gene expression. J. Biol. Chem., 284, 28729–28737.
15.
KangM., PiliszekA., ArtusJ., and HadjantonakisA.K. (2013). FGF4 is required for lineage restriction and salt-and-pepper distribution of primitive endoderm factors but not their initial expression in the mouse. Development, 140, 267–279.
16.
KirchhofN., CarnwathJ.W., LemmeE., AnastassiadisK., ScholerH., and NiemannH. (2000). Expression pattern of Oct-4 in preimplantation embryos of different species. Biol. Reprod., 63, 1698–1705.
17.
KuijkE.W., Du PuyL., Van TolH.T., OeiC.H., HaagsmanH.P., ColenbranderB., and RoelenB.A. (2008). Differences in early lineage segregation between mammals. Dev. Dyn., 237, 918–927.
18.
KurodaT., TadaM., KubotaH., KimuraH., HatanoS.Y., 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.
19.
KurosakaS., EckardtS., and McLaughlinK.J. (2004). Pluripotent lineage definition in bovine embryos by Oct4 transcript localization. Biol. Reprod., 71, 1578–1582.
20.
Le BinG.C., Munoz-DescalzoS., KurowskiA., LeitchH., LouX., MansfieldW., Etienne-DumeauC., GraboleN., MulasC., NiwaH., et al. (2014). Oct4 is required for lineage priming in the developing inner cell mass of the mouse blastocyst. Development, 141, 1001–1010.
21.
MaY.G., RosfjordE., HuebertC., WilderP., TiesmanJ., KellyD., and RizzinoA. (1992). Transcriptional regulation of the murine k-FGF gene in embryonic cell lines. Dev. Biol., 154, 45–54.
22.
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.
23.
NagatomoH., KagawaS., KishiY., TakumaT., SadaA., YamanakaK., AbeY., WadaY., TakahashiM., KonoT.et al. (2013). Transcriptional wiring for establishing cell lineage specification at the blastocyst stage in cattle. Biol. Reprod., 88, 158.
24.
NganvongpanitK., MullerH., RingsF., HoelkerM., JennenD., TholenE., HavlicekV., BesenfelderU., SchellanderK., and TesfayeD. (2006). Selective degradation of maternal and embryonic transcripts in in vitro produced bovine oocytes and embryos using sequence specific double-stranded RNA. Reproduction, 131, 861–874.
25.
NicholsJ., ZevnikB., AnastassiadisK., NiwaH., Klewe-NebeniusD., ChambersI., ScholerH., and SmithA. (1998). Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell, 95, 379–391.
26.
NishiokaN., InoueK., AdachiK., KiyonariH., OtaM., RalstonA., YabutaN., HiraharaS., StephensonR.O., OgonukiN., et al. (2009). The Hippo signaling pathway components Lats and Yap pattern Tead4 activity to distinguish mouse trophectoderm from inner cell mass. Dev. Cell, 16, 398–410.
27.
NishiokaN., YamamotoS., KiyonariH., SatoH., SawadaA., OtaM., NakaoK., and SasakiH. (2008). Tead4 is required for specification of trophectoderm in pre-implantation mouse embryos. Mech. Dev., 125, 270–283.
28.
NiswanderL., and MartinG.R. (1992). Fgf-4 expression during gastrulation, myogenesis, limb and tooth development in the mouse. Development, 114, 755–768.
29.
NiwaH., MasuiS., ChambersI., SmithA.G., and MiyazakiJ. (2002). Phenotypic complementation establishes requirements for specific POU domain and generic transactivation function of Oct-3/4 in embryonic stem cells. Mol. Cell. Biol., 22, 1526–1536.
30.
NiwaH., MiyazakiJ., and SmithA.G. (2000). Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nat. Genet., 24, 372–376.
31.
NiwaH., ToyookaY., ShimosatoD., StrumpfD., TakahashiK., YagiR., and RossantJ. (2005). Interaction between Oct3/4 and Cdx2 determines trophectoderm differentiation. Cell, 123, 917–929.
32.
PalmieriS.L., PeterW., HessH., and ScholerH.R. (1994). Oct-4 transcription factor is differentially expressed in the mouse embryo during establishment of the first two extraembryonic cell lineages involved in implantation. Dev. Biol., 166, 259–267.
33.
PedersenR.A., WuK., and BalakierH. (1986). Origin of the inner cell mass in mouse embryos: cell lineage analysis by microinjection. Dev. Biol., 117, 581–595.
34.
RalstonA., CoxB.J., NishiokaN., SasakiH., CheaE., Rugg-GunnP., GuoG., RobsonP., DraperJ.S., and RossantJ. (2010). Gata3 regulates trophoblast development downstream of Tead4 and in parallel to Cdx2. Development, 137, 395–403.
35.
RalstonA., and RossantJ. (2008). Cdx2 acts downstream of cell polarization to cell-autonomously promote trophectoderm fate in the early mouse embryo. Dev. Biol., 313, 614–629.
36.
RappoleeD.A., BasilicoC., PatelY., and WerbZ. (1994). Expression and function of FGF-4 in peri-implantation development in mouse embryos. Development, 120, 2259–2269.
37.
RoddaD.J., ChewJ.L., LimL.H., LohY.H., WangB., NgH.H., and RobsonP. (2005). Transcriptional regulation of Nanog by OCT4 and SOX2. J. Biol. Chem., 280, 24731–24737.
38.
RossantJ., and TamP.P. (2009). Blastocyst lineage formation, early embryonic asymmetries and axis patterning in the mouse. Development, 136, 701–713.
39.
SakuraiN., FujiiT., HashizumeT., and SawaiK. (2013). Effects of downregulating oct-4 transcript by RNA interference on early development of porcine embryos. J. Reprod. Dev., 59, 353–360.
40.
SawaiK., FujiiT., HirayamaH., HashizumeT., and MinamihashiA. (2012). Epigenetic status and full-term development of bovine cloned embryos treated with trichostatin A. J. Reprod. Dev., 58, 302–309.
41.
SchiffmacherA.T., and KeeferC.L. (2013). CDX2 regulates multiple trophoblast genes in bovine trophectoderm CT-1 cells. Mol. Reprod. Dev., 80, 826–839.
42.
SchlessingerJ., PlotnikovA.N., IbrahimiO.A., EliseenkovaA.V., YehB.K., YayonA., LinhardtR.J., and MohammadiM. (2000). Crystal structure of a ternary FGF-FGFR-heparin complex reveals a dual role for heparin in FGFR binding and dimerization. Mol. Cell, 6, 743–750.
43.
StrumpfD., MaoC.A., YamanakaY., RalstonA., ChawengsaksophakK., BeckF., and RossantJ. (2005). Cdx2 is required for correct cell fate specification and differentiation of trophectoderm in the mouse blastocyst. Development, 132, 2093–2102.
44.
TakahashiK., and YamanakaS. (2006). Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell, 126, 663–676.
45.
TanakaS., KunathT., HadjantonakisA.K., NagyA., and RossantJ. (1998). Promotion of trophoblast stem cell proliferation by FGF4. Science, 282, 2072–2075.
46.
van EijkM.J., van RooijenM.A., ModinaS., ScesiL., FolkersG., van TolH.T., BeversM.M., FisherS.R., LewinH.A., RakacolliD., et al. (1999). Molecular cloning, genetic mapping, and developmental expression of bovine POU5F1. Biol. Reprod., 60, 1093–1103.
47.
WuG., GentileL., FuchikamiT., SutterJ., PsathakiK., EstevesT.C., Arauzo-BravoM.J., OrtmeierC., VerberkG., AbeK.et al. (2010). Initiation of trophectoderm lineage specification in mouse embryos is independent of Cdx2. Development, 137, 4159–4169.
48.
YagiR., KohnM.J., KaravanovaI., KanekoK.J., VullhorstD., DePamphilisM.L., and BuonannoA. (2007). Transcription factor TEAD4 specifies the trophectoderm lineage at the beginning of mammalian development. Development, 134, 3827–3836.
49.
YamanakaY., LannerF., and RossantJ. (2010). FGF signal-dependent segregation of primitive endoderm and epiblast in the mouse blastocyst. Development, 137, 715–724.
50.
YamashitaS., AbeH., ItohT., SatohT., and HoshiH. (1999). A serum-free culture system for efficient in vitro production of bovine blastocysts with improved viability after freezing and thawing. Cytotechnology, 31, 123–131.
51.
YuanH., CorbiN., BasilicoC., and DaileyL. (1995). Developmental-specific activity of the FGF-4 enhancer requires the synergistic action of Sox2 and Oct-3. Genes Dev. 9, 2635–2645.
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