Rex1/Zfp42 is a nuclear protein that is highly conserved in mammals, and widely used as an embryonic stem (ES) cell marker. Although Rex1 expression is associated with enhanced pluripotency, loss-of-function models recently described do not exhibit major phenotypes, and both preimplantation development and ES cell derivation appear normal in the absence of Rex1. To better understand the functional role of Rex1, we examined the expression and localization of Rex1 during preimplantation development. Our studies indicated that REX1 is expressed at all stages during mouse preimplantation development, with a mixed pattern of nuclear, perinuclear, and cytoplasmic localization. Chromatin association seemed to be altered in 8-cell embryos, and in the blastocyst, we found REX1 localized almost exclusively in the nucleus. A functional role for Rex1 in vivo was assessed by gain- and loss-of-function approaches. Embryos with attenuated levels of Rex1 after injection of zygotes with siRNAs did not exhibit defects in preimplantation development in vitro. In contrast, overexpression of Rex1 interfered with cleavage divisions and with proper blastocyst development, although we failed to detect alterations in the expression of lineage and pluripotency markers. Rex1 gain- and loss-of-function did alter the expression levels of Zscan4, an important regulator of preimplantation development and pluripotency. Our results suggest that Rex1 plays a role during preimplantation development. They are compatible with a role for Rex1 during acquisition of pluripotency in the blastocyst.
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References
1.
EdwardsRG. 2003. Aspects of the molecular regulation of early mammalian development. Reprod Biomed Online, 6:97–113.
2.
RossantJ. 2001. Stem cells from the mammalian blastocyst. Stem Cells, 19:477–482.
3.
JohnsonMH, McConnellJM. 2004. Lineage allocation and cell polarity during mouse embryogenesis. Semin Cell Dev Biol, 15:583–597.
4.
HamataniT, CarterMG, SharovAA, KoMS. 2004. Dynamics of global gene expression changes during mouse preimplantation development. Dev Cell, 6:117–131.
5.
EvansMJ, KaufmanMH. 1981. Establishment in culture of pluripotential cells from mouse embryos. Nature, 292:154–156.
6.
MartinGR. 1981. Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc Natl Acad Sci U S A, 78:7634–7638.
7.
NicholsJ, ZevnikB, AnastassiadisK, NiwaH, Klewe-NebeniusD, ChambersI, SchölerH, SmithA. 1998. Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell, 95:379–391.
8.
AvilionA, NicolisS, PevnyL, PerezL, VivianN, Lovell-BadgeR. 2003. Multipotent cell lineages in early mouse development depend on SOX2 function. Genes Dev, 17:126–140.
9.
ChambersI, ColbyD, RobertsonM, NicholsJ, LeeS, TweedieS, SmithA. 2003. Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell, 113:643–655.
10.
MitsuiK, TokuzawaY, ItohH, SegawaK, MurakamiM, TakahashiK, MaruyamaM, MaedaM, YamanakaS. 2003. The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell, 113:631–642.
11.
TakahashiK, YamanakaS. 2006. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell, 126:663–676.
12.
ChenX, XuH, YuanP, FangF, HussM, VegaVB, WongE, OrlovYL, ZhangWet al.2008. Integration of external signaling pathways with the core transcriptional network in embryonic stem cells. Cell, 133:1106–1117.
13.
YoshikawaT, PiaoY, ZhongJ, MatobaR, CarterMG, WangY, GoldbergI, KoMS. 2006. High-throughput screen for genes predominantly expressed in the ICM of mouse blastocysts by whole mount in situ hybridization. Gene Expr Patterns, 6:213–224.
14.
ZhangJ, GaoW, YangH, ZhangB, ZhuZ, XueY. 2006. Screening for genes essential for mouse embryonic stem cell self-renewal using a subtractive RNA interference library. Stem Cells, 24:2661–2668.
15.
HuG, KimJ, XuQ, LengY, OrkinSH, ElledgeSJ. 2009. A genome-wide RNAi screen identifies a new transcriptional module required for self-renewal. Genes Dev, 23:837–848.
ZhangW, WalkerE, TamplinOJ, RossantJ, StanfordWL, HughesTR. 2006. Zfp206 regulates ES cell gene expression and differentiation. Nucleic Acids Res, 34:4780–4790.
18.
FalcoG, LeeSL, StanghelliniI, BasseyUC, HamataniT, KoMS. 2007. Zscan4: a novel gene expressed exclusively in late 2-cell embryos and embryonic stem cells. Dev Biol, 307:539–550.
19.
CarterMG, StaggCA, FalcoG, YoshikawaT, BasseyUC, AibaK, SharovaLV, ShaikN, KoMS. 2008. An in situ hybridization-based screen for heterogeneously expressed genes in mouse ES cells. Gene Expr Patterns, 8:181–198.
20.
ZalzmanM, FalcoG, SharovaLV, NishiyamaA, ThomasM, LeeSL, StaggCA, HoangHG, YangHTet al.2010. Zscan4 regulates telomere elongation and genomic stability in ES cells. Nature, 464:858–863.
21.
KimJ, FaulkC, KimJ. 2007. Retroposition and evolution of the DNA-binding motifs of YY1, YY2 and REX1. Nucleic Acids Res, 35:3442–3452.
22.
HoslerB, LaRosaG, GrippoJ, GudasL. 1989. Expression of REX-1, a gene containing zinc finger motifs, is rapidly reduced by retinoic acid in F9 teratocarcinoma cells. Mol Cell Biol, 9:5623–5629.
23.
RogersM, HoslerB, GudasL. 1991. Specific expression of a retinoic acid-regulated, zinc-finger gene, Rex-1, in preimplantation embryos, trophoblast and spermatocytes. Development, 113:815–824.
KarlmarkK, FreilingerA, MartonE, RosnerM, LubecG, HengstschlägerM. 2005. Activation of ectopic Oct-4 and Rex-1 promoters in human amniotic fluid cells. Int J Mol Med, 16:987–992.
26.
KristensenDM, NielsenJE, SkakkebaekNE, GraemN, JacobsenGK, Rajpert-De MeytsE, LeffersH. 2008. Presumed pluripotency markers UTF-1 and REX-1 are expressed in human adult testes and germ cell neoplasms. Hum Reprod, 23:775–782.
ToyookaY, ShimosatoD, MurakamiK, TakahashiK, NiwaH. 2008. Identification and characterization of subpopulations in undifferentiated ES cell culture. Development, 135:909–918.
29.
BrivanlouA, GageF, JaenischR, JessellT, MeltonD, RossantJ. 2003. Stem cells. Setting standards for human embryonic stem cells. Science, 300:913–916.
30.
ChanE, RatanasirintrawootS, ParkI, ManosP, LohY, HuoH, MillerJ, HartungO, RhoJet al.2009. Live cell imaging distinguishes bona fide human iPS cells from partially reprogrammed cells. Nat Biotechnol, 27:1033–1037.
31.
ScotlandK, ChenS, SylvesterR, GudasL. 2009. Analysis of Rex1 (zfp42) function in embryonic stem cell differentiation. Dev Dyn, 238:1863–1877.
32.
MasuiS, OhtsukaS, YagiR, TakahashiK, KoM, NiwaH. 2008. Rex1/Zfp42 is dispensable for pluripotency in mouse ES cells. BMC Dev Biol, 8:45.
33.
Garcia-TuñonI, GuallarD, Alonso-MartinS, BenitoAA, Benítez-LázaroA, Pérez-PalaciosR, MuniesaP, ClimentM, SánchezM, VidalM, SchoorlemmerJ. 2011. Association of Rex-1 to target genes supports its interaction with polycomb function. Stem Cell Res, 7:1–16.
34.
KimJD, KimH, EkramMB, YuS, FaulkC, KimJ. 2011. Rex1/Zfp42 as an epigenetic regulator for genomic imprinting. Hum Mol Genet, 20:1353–1362.
35.
RezendeNC, LeeMY, MonetteS, MarkW, LuA, GudasLJ. 2011. Rex1 (Zfp42) null mice show impaired testicular function, abnormal testis morphology, and aberrant gene expression. Dev Biol, 356:370–382.
36.
GuallarD, Pérez-PalaciosR, ClimentM, Martínez-AbadíaI, LarragaA, Fernández-JuanM, VallejoC, MuniesaP, SchoorlemmerJ. 2012. Expression of endogenous retroviruses is negatively regulated by the pluripotency marker Rex1/Zfp42. Nucleic Acids Res, 2012[Epub ahead of print]10.1093/nar/gks686.
37.
Torres-PadillaM, BannisterA, HurdP, KouzaridesT, Zernicka-GoetzM. 2006. Dynamic distribution of the replacement histone variant H3.3 in the mouse oocyte and preimplantation embryos. Int J Dev Biol, 50:455–461.
38.
NiwaH, BurdonT, ChambersI, SmithA. 1998. Self-renewal of pluripotent embryonic stem cells is mediated via activation of STAT3. Genes Dev, 12:2048–2060.
39.
NagyA, GertsensteinM, VinterstenK, BehringerR. 2003. Manipulating the Mouse Embryo: A Laboratory Manual. Cold Spring Harbor Laboratory Press: Cold Spring Harbor, NY.
40.
ZengF, BaldwinDA, SchultzRM. 2004. Transcript profiling during preimplantation mouse development. Dev Biol, 272:483–496.
41.
Ruddock-D'CruzN, PrashadkumarS, WilsonK, HeffernanC, CooneyM, FrenchA, JansD, VermaP, HollandM. 2008. Dynamic changes in localization of Chromobox (Cbx) family members during the maternal to embryonic transition. Mol Reprod Dev, 75:477–488.
42.
WorradDM, RamPT, SchultzRM. 1994. Regulation of gene expression in the mouse oocyte and early preimplantation embryo: developmental changes in Sp1 and TATA box-binding protein, TBP. Development, 120:2347–2357.
43.
DonohoeM, ZhangX, McGinnisL, BiggersJ, LiE, ShiY. 1999. Targeted disruption of mouse Yin Yang 1 transcription factor results in peri-implantation lethality. Mol Cell Biol, 19:7237–7244.
44.
NavarroP, OldfieldA, LegoupiJ, FestucciaN, DuboisA, AttiaM, SchoorlemmerJ, RougeulleC, ChambersI, AvnerP. 2010. Molecular coupling of Tsix regulation and pluripotency. Nature, 468:457–460.
45.
MiyanariY, Torres-PadillaME. 2012. Control of ground-state pluripotency by allelic regulation of Nanog. Nature, 483:470–473.
46.
MacfarlanTS, GiffordWD, AgarwalS, DriscollS, LettieriK, WangJ, AndrewsSE, FrancoL, RosenfeldMG, RenB, PfaffSL. 2011. Endogenous retroviruses and neighboring genes are coordinately repressed by LSD1/KDM1A. Genes Dev, 25:594–607.
47.
PeastonAE, EvsikovAV, GraberJH, de VriesWN, HolbrookAE, SolterD, KnowlesBB. 2004. Retrotransposons regulate host genes in mouse oocytes and preimplantation embryos. Dev Cell, 7:597–606.
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