Primordial germ cells (PGCs), the precursors of gametes, have regulatory mechanisms involving transcription factors and epigenetic modifications. The transcription factor NANOG is a key regulator of germ cell and embryonic stem cell characteristics. However, the epigenetic regulation of NANOG with a histone deacetylase (HDAC) complex in PGCs has not been studied. In this study, we investigated the epigenetic regulation, and in particular the histone acetylation, of NANOG in chicken PGCs. Intriguingly, although NANOG was highly activated in chicken PGCs, the upstream region of its promoter was moderately suppressed by histone deacetylation. HDAC inhibition induced histone H3 lysine 9 acetylation (H3K9ac) and derepressed NANOG expression. Furthermore, knockdown studies revealed that HDAC complex members, such as RE1-silencing transcription factor (REST) and REST corepressor 3 (RCOR3), are important epigenetic modulators of NANOG expression in chicken PGCs. We demonstrate that moderate regulation of NANOG by the REST/CoREST/HDAC complex might be crucial for maintaining the integrity of PGCs.
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References
1.
TangWW, KobayashiT, IrieN, DietmannS and SuraniMA. (2016). Specification and epigenetic programming of the human germ line. Nat Rev Genet, 17:585–600.
2.
StromeS and UpdikeD. (2015). Specifying and protecting germ cell fate. Nat Rev Mol Cell Biol, 16:406–416.
3.
SekiY, HayashiK, ItohK, MizugakiM, SaitouM and MatsuiY. (2005). Extensive and orderly reprogramming of genome-wide chromatin modifications associated with specification and early development of germ cells in mice. Dev Biol, 278:440–458.
4.
AkiyamaT, NagataM and AokiF. (2006). Inadequate histone deacetylation during oocyte meiosis causes aneuploidy and embryo death in mice. Proc Natl Acad Sci U S A, 103:7339–7344.
5.
SteilmannC, ParadowskaA, BartkuhnM, ViewegM, SchuppeHC, BergmannM, KlieschS, WeidnerW and StegerK. (2011). Presence of histone H3 acetylated at lysine 9 in male germ cells and its distribution pattern in the genome of human spermatozoa. Reprod Fertil Dev, 23:997–1011.
6.
HanJY. (2009). Germ cells and transgenesis in chickens. Comp Immunol Microbiol Infect Dis, 32:61–80.
7.
TsunekawaN, NaitoM, SakaiY, NishidaT and NoceT. (2000). Isolation of chicken vasa homolog gene and tracing the origin of primordial germ cells. Development, 127:2741–2750.
8.
LeeHC, ChoiHJ, LeeHG, LimJM, OnoT and HanJY. (2016). DAZL expression explains origin and central formation of primordial germ cells in chickens. Stem Cells Dev, 25:68–79.
9.
HamburgerV and HamiltonHL. (1951). A series of normal stages in the development of the chick embryo. J Morphol, 88:49–92.
10.
KressC, MontilletG, JeanC, FuetA and PainB. (2016). Chicken embryonic stem cells and primordial germ cells display different heterochromatic histone marks than their mammalian counterparts. Epigenetics Chromatin, 9:5.
11.
YuJ, VodyanikMA, Smuga-OttoK, Antosiewicz-BourgetJ, FraneJL, TianS, NieJ, JonsdottirGA, RuottiV, et al. (2007). Induced pluripotent stem cell lines derived from human somatic cells. Science, 318:1917–1920.
12.
YamaguchiS, KimuraH, TadaM, NakatsujiN and TadaT. (2005). Nanog expression in mouse germ cell development. Gene Expr Patterns, 5:639–646.
13.
IrieN, TangWW and Azim SuraniM. (2014). Germ cell specification and pluripotency in mammals: a perspective from early embryogenesis. Reprod Med Biol, 13:203–215.
14.
YamaguchiS, KurimotoK, YabutaY, SasakiH, NakatsujiN, SaitouM and TadaT. (2009). Conditional knockdown of Nanog induces apoptotic cell death in mouse migrating primordial germ cells. Development, 136:4011–4020.
WesternPS, van den BergenJA, MilesDC and SinclairAH. (2010). Male fetal germ cell differentiation involves complex repression of the regulatory network controlling pluripotency. FASEB J, 24:3026–3035.
17.
ZhangM, LeitchHG, TangWWC, FestucciaN, Hall-PonseleE, NicholsJ, SuraniMA, SmithA and ChambersI. (2018). Esrrb complementation rescues development of Nanog-null germ cells. Cell Rep, 22:332–339.
18.
LavialF, AcloqueH, BertocchiniF, MacleodDJ, BoastS, BachelardE, MontilletG, ThenotS, SangHM, et al. (2007). The Oct4 homologue PouV and Nanog regulate pluripotency in chicken embryonic stem cells. Development, 134:3549–3563.
19.
HanJY, LeeHG, ParkYH, HwangYS, KimSK, RengarajD, ChoBW and LimJM. (2018). Acquisition of pluripotency in the chick embryo occurs during intrauterine embryonic development via a unique transcriptional network. J Anim Sci Biotechnol, 9:31.
20.
NakanohS, FuseN, TadokoroR, TakahashiY and AgataK. (2017). Jak1/Stat3 signaling acts as a positive regulator of pluripotency in chicken pre-gastrula embryos. Dev Biol, 421:43–51.
21.
CañónS, HerranzC and ManzanaresM. (2006). Germ cell restricted expression of chick Nanog. Dev Dyn, 235:2889–2894.
22.
MurakamiK, GünesdoganU, ZyliczJJ, TangWWC, SenguptaR, KobayashiT, KimS, ButlerR, DietmannS and SuraniMA. (2016). NANOG alone induces germ cells in primed epiblast in vitro by activation of enhancers. Nature, 529:403–407.
23.
YangP, WangY, ChenJ, LiH, KangL, ZhangY, ChenS, ZhuB and GaoS. (2011). RCOR2 is a subunit of the LSD1 complex that regulates ESC property and substitutes for SOX2 in reprogramming somatic cells to pluripotency. Stem Cells, 29:791–801.
24.
dos SantosRL, TostiL, RadzisheuskayaA, CaballeroIM, KajiK, HendrichB and SilvaJC. (2014). MBD3/NuRD facilitates induction of pluripotency in a context-dependent manner. Cell Stem Cell, 15:102–110.
25.
SaundersA, HuangX, FidalgoM, ReimerMHJr., FaiolaF, DingJ, Sánchez-PriegoC, GuallarD, SáenzC, LiD and WangJ. (2017). The SIN3A/HDAC corepressor complex functionally cooperates with NANOG to promote pluripotency. Cell Rep, 18:1713–1726.
26.
ParkTS and HanJY. (2012). piggyBac transposition into primordial germ cells is an efficient tool for transgenesis in chickens. Proc Natl Acad Sci U S A, 109:9337–9341.
27.
ParkTS, LeeHJ, KimKH, KimJS and HanJY. (2014). Targeted gene knockout in chickens mediated by TALENs. Proc Natl Acad Sci U S A, 111:12716–12721.
28.
KoMH, HwangYS, RimJS, HanHJ and HanJY. (2017). Avian blastoderm dormancy arrests cells in G2 and suppresses apoptosis. FASEB J, 31:3240–3250.
29.
KazantsevAG and ThompsonLM. (2008). Therapeutic application of histone deacetylase inhibitors for central nervous system disorders. Nat Rev Drug Discov, 7:854–868.
30.
KwonHJ, KimMS, KimMJ, NakajimaH and KimKW. (2002). Histone deacetylase inhibitor FK228 inhibits tumor angiogenesis. Int J Cancer, 97:290–296.
31.
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.
32.
LohYH, WuQ, ChewJL, VegaVB, ZhangW, ChenX, BourqueG, GeorgeJ, LeongB, et al. (2006). The Oct4 and Nanog transcription network regulates pluripotency in mouse embryonic stem cells. Nat Genet, 38:431–440.
33.
WareCB, WangL, MechamBH, ShenL, NelsonAM, BarM, LambaDA, DauphinDS, BuckinghamB, et al. (2009). Histone deacetylase inhibition elicits an evolutionarily conserved self-renewal program in embryonic stem cells. Cell Stem Cell, 4:359–369.
HartAH, HartleyL, ParkerK, IbrahimM, LooijengaLH, PauchnikM, ChowCW and RobbL. (2005). The pluripotency homeobox gene NANOG is expressed in human germ cell tumors. Cancer, 104:2092–2098.
36.
HeaneyJD, AndersonEL, MichelsonMV, ZechelJL, ConradPA, PageDC and NadeauJH. (2012). Germ cell pluripotency, premature differentiation and susceptibility to testicular teratomas in mice. Development, 139:1577–1586.
37.
JuricD, SaleS, HromasRA, YuR, WangY, DuranGE, TibshiraniR, EinhornLH and SikicBI. (2005). Gene expression profiling differentiates germ cell tumors from other cancers and defines subtype-specific signatures. Proc Natl Acad Sci U S A, 102:17763–17768.
38.
SteblerJ, SpielerD, SlanchevK, MolyneauxKA, RichterU, CojocaruV, TarabykinV, WylieC, KesselM and RazE. (2004). Primordial germ cell migration in the chick and mouse embryo: the role of the chemokine SDF-1/CXCL12. Dev Biol, 272:351–361.
39.
SchneiderDT, SchusterAE, FritschMK, HuJ, OlsonT, LauerS, GöbelU and PerlmanEJ. (2001). Multipoint imprinting analysis indicates a common precursor cell for gonadal and nongonadal pediatric germ cell tumors. Cancer Res, 61:7268–7276.
40.
ChongJA, Tapia-RamírezJ, KimS, Toledo-AralJJ, ZhengY, BoutrosMC, AltshullerYM, FrohmanMA, KranerSD and MandelG. (1995). REST: a mammalian silencer protein that restricts sodium channel gene expression to neurons. Cell, 80:949–957.
41.
ChenZF, PaquetteAJ and AndersonDJ. (1998). NRSF/REST is required in vivo for repression of multiple neuronal target genes during embryogenesis. Nat Genet, 20:136–142.
42.
GrimesJA, NielsenSJ, BattaglioliE, MiskaEA, SpehJC, BerryDL, AtoufF, HoldenerBC, MandelG and KouzaridesT. (2000). The co-repressor mSin3A is a functional component of the REST-CoREST repressor complex. J Biol Chem, 275:9461–9467.
43.
JohnsonR, TehCH, KunarsoG, WongKY, SrinivasanG, CooperML, VoltaM, ChanSS, LipovichL, et al. (2008). REST regulates distinct transcriptional networks in embryonic and neural stem cells. PLoS Biol, 6:e256.
44.
YuHB, JohnsonR, KunarsoG and StantonLW. (2011). Coassembly of REST and its cofactors at sites of gene repression in embryonic stem cells. Genome Res, 21:1284–1293.
45.
LeeM, JiH, FurutaY, ParkJI and McCreaPD. (2014). p120-catenin regulates REST and CoREST, and modulates mouse embryonic stem cell differentiation. J Cell Sci, 127:4037–4051.
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