Epigenetic reprogramming, gene expression and in vitro development of porcine SCNT embryos are significantly improved by a histone deacetylase inhibitor—m-carboxycinnamic acid bishydroxamide (CBHA)

Abstract

Insufficient epigenetic reprogramming of donor nuclei is believed to be one of the most important causes of low development efficiency of mammalian somatic cell nuclear transfer (SCNT). Previous studies have shown that both the in vitro and in vivo development of mouse SCNT embryos could be increased significantly by treatment with various histone deacetylase inhibitors (HDACi), including Trichostatin A, Scriptaid, and m-carboxycinnamic acid bishydroxamide (CBHA), in which only the effect of CBHA has not yet been tested in other species. In this paperweexamine the effect ofCBHAtreatment on the development of porcine SCNT embryos. We have discovered the optimum dosage and time for CBHA treatment: incubating SCNT embryos with 2 μmol/L CBHA for 24 h after activation could increase the blastocyst rate from 12.7% to 26.5%. Immunofluorescence results showed that the level of acetylation at histone 3 lysine 9 (AcH3K9), acetylation at histone 3 lysine 18 (AcH3K18), and acetylation at histone 4 lysine 16 (AcH4K16) was raised after CBHAtreatment. Meanwhile,CBHAtreatment improved the expression of development relating genes such as pou5f1, cdx2, and the imprinted genes like igf2. Despite these promising in vitro results and histone reprogramming, the full term development was not significantly increased after treatment. In conclusion, CBHA improves the in vitro development of pig SCNT embryos, increases the global histone acetylation and corrects the expression of some developmentally important genes at early stages. As in mouse SCNT, we have shown that nuclear epigenetic reprogramming in pig early SCNTembryos can be modified by CBHA treatment.

Keywords

swine / nuclear transfer / epigenetic reprogramming / histone deacetylase inhibitor

References

[1] Baguisi A, Behboodi E, Melican DT, Pollock JS, Destrempes MM (1999) Production of goats by somatic cell nuclear transfer. Nat Biotechnol17: 456-461

[2] Bendixen E, Danielsen M, Larsen K, Bendixen C (2013) Advances in porcine genomics and proteomics-a toolbox for developing the pig as a model organism for molecular biomedical research. Brief Funct Genomics9(3): 208-219

[3] Bock C, Kiskinis E, Verstappen G (2011) Reference maps of human ES and iPS cell variation enable high-throughput characterization of pluripotent cell lines. Cell144: 439-452

[4] Chawengsaksophac K, Graaff W, Beck F (2004) Cdx2 is essential for axial elongation in mouse development. PNAS101: 7641-7645

[5] Chesne P, Adenot PG, Viglietta C, Baratte M, Boulanger L (2002) Cloned rabbits produced by nuclear transfer from adult somatic cells. Nat Biotechnol20: 366-369

[6] Dai XP, Hao J, Hou XJ, Hai T, Zhou Q (2010) Somatic nucleus reprogramming is signiflcantly improved by m-carboxycinnamic acid bishydroxamide, a histone deacetylase inhibitor. J Biol Chem285: 31002-31010

[7] Deshmukh RS, Ostrup O, Hyttel P (2012) Early aberrations in chromatin dynamics in embryos produced under in vitro conditions. Cell Reprogram14: 225-234

[8] Ekser B, Ezzelarab M, Hara H, Windt D, Wijkstrom M, Bottino R, Trucco M, Cooper DKC (2012) Clinical xenotransplantation: the next medical revolution? Lancet379: 672-683

[9] Galli C, Lagutina I, Crotti G, Colleoni S, Turini P (2003) A cloned horse born to its dam twin (vol 424, p 635, 2003). Nature425: 680

[10] Gock H, Nottle M, Lew AM, d’Apice AJ, Cowan P (2011) Genetic modiflcation of pigs for solid organ xenotransplantation. Transplant Rev25: 9-20

[11] Han YM, Kang YK, Koo DB, Lee KK (2003) Nuclear reprogramming of cloned embryos produced in vitro. Theriogenology59: 33-34

[12] Kato Y, Tani T, Sotomaru Y, Kurokawa K, Kato J (1998) Eight calves cloned from somatic cells of a single adult. Science282: 2095-2098

[13] Kim YJ, Ahn KS, Kim M, Shim H (2011) Comparison of potency between histone deacetylase inhibitors trichostatin A and valproic acid on enhancing in vitro development f porcine somatic cell nuclear transfer embryos. In Vitro Cell Dev Biol Anim47: 283-289

[14] Kishigami S, Mizutani E, Ohta H (2006) Signiflcant improvement of mouse cloning technique by treatment with trichostain A after somatic nuclear transfer. Biochem Biophys Res Commun340: 183-189

[15] Kumar BM, Jin HF, Kim JG, Ock SA, Hong Y, Balasubramanian S (2007) Differential gene expression patterns in porcine nuclear transfer embryos reconstructed with fetal flbroblasts and mesenchymal stem cells. Dev Dyn236: 435-446

[16] Lager AE, Ragina NP, Cibelli JB (2008) Trichostatin A improves histone acetylation in bovine somatic cell nuclear transfer early embryos. Cloning Stem Cells10: 371-379

[17] Latham KE, Doherty AS, Scott CD, Schultz RM (1994) Igf2r and Igf2 gene expression in androgenetic, gynogenetic, and parthenogenetic preimplantation mouse embryos: absence of regulation by genomic imprinting. Genes Dev8: 290-299

[18] Lee JH, Hart SRL, Skalnik DG (2004) Histone deacetylase activity is required for embryonic stem cell differentiation. Genesis38: 32-38

[19] Lee BC, Kim MK, Jang G, Oh HJ, Yuda F (2005) Dogs cloned from adult somatic cells. Nature436: 641

[20] Lee E, Lee SH, Lee BC (2006) Analysis of nuclear reprogramming in cloned miniature pig embryos by expression of Oct-4 and Oct-4 related genes. Biochem Biophys Res Commun348: 1419-1428

[21] Li E (2002) Chromatin modiflcation and epigenetic reprogramming in mammalian development. Nature3: 662-673

[22] Li ZY, Sun XS, Chen J, Liu XM, Wisely SM (2006) Cloned ferrets produced by somatic cell nuclear transfer. Dev Biol293: 439-448

[23] Li J, Svarcova O, Villemoes K, Vajta G (2008) High in vitro development after somatic cell nuclear transfer and trichostatin A treatment of reconstructed porcine embryos. Theriogenology70: 800-808

[24] Loh YH, Wu Q, Ng HH (2006) The Oct4 and Nanog transcription network regulated pluripotency in mouse embryonic stem cells. Nat Genet38: 431-440

[25] Luo YL, Lin L, Bolund L, Jesen TG, Sorensen CB (2012) Genetically modifled pigs for biomedical research. J Inherit Metab Dis35: 695-713

[26] Maalouf W, Liu ZC, Zink D (2009) Trichostatin A treatment of cloned mouse embryos improves constitutive heterochromatin remodeling as well as developmental potential to term. BMC Dev Biol9: 11

[27] Mason K, Liu ZC, Beaujean N (2012) Chromatin and epigenetic modiflcations during early mammalian development. Anim Reprod Sci134: 45-55

[28] Mattout A, Biran A, Meshorer E (2011) Global epigenetic changes during somatic cell reprogramming to iPS cells. J Mol Biol3: 341-350

[29] Menendez S, Camus S, Herreria A (2011) Increased dosage of tumor suppressors limits the tumorigenicity of cells without affecting their pluripotency. Aging Cell11: 41-50

[30] Miyoshi K, Mori H, Mizobe Y, Sato M (2010) Beneflcal effects of reversine on in vitro development of miniature pig somatic cell nuclear transfer embryos. J Reprod Dev56: 291-296

[31] Monteriro FM, Oliveira CS, Oliveira LZ, Garcia JM (2011) Chromatin modifying agents in the vitro production of bovine embryos. SAGE-Hindawi access to research veterinary medicine international

[32] Niwa H, Toyooka Y, Rossant J (2005) Interaction between Oct3/ 4 and Cdx2 determines trophectodern differentiation. Cell123: 917-929

[33] Ogura A, Inoue K, Wakayama T (2013) Recent advancements in cloning by somatic cell nuclear transfer. Philos Trans R Soc368: 20110329

[34] Okita K, Ichisaka T, Yamanaka S (2007) Generation of germline-competent induced pluripotent stem cells. Nature448: 313-318

[35] Onishi A, Iwamoto M, Akita T, Mikawa S, Takeda K (2000) Pig cloning by microinjection of fetal flbroblast nuclei. Science289: 1188-1190

[36] Pesce M, Scholer HR (2001) Oct-4: gatekeeper in the beginnings of mammalian development. Stem Cells19: 271-278

[37] Peserico A, Simone C (2011) Physical and functional HAT/ HDAC interplay regulates protein acetylation balance. J Biomed Biotechnol2011: 371832

[38] Polejaeva IA, Chen SH, Vaught TD, Page RL, Mullins J, Ball S, Dal Y, Boone J (2000) Cloned pigs produced by nuclear transfer from adult somatic cells. Nature407: 86-90

[39] Rybouchkin A, Kato Y, Tsunoda Y (2006) Role of histone acetylation in reprogramming of somatic nuclei following nuclear transfer. Biol Reprod74: 1083-1089

[40] Shin T, Kraemer D, Pryor J, Liu L, Rugila J (2002) A cat cloned by nuclear transplantation. Nature415: 859

[41] Strumpf D, Mao CA, Yamanaka Y, Rossant J (2005) Cdx2 is required for correct cell fate speciflcation and differentiation of trophectoderm in the mouse blastocyst. Development132: 2093-2102

[42] Takahashi K, Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult flbroblast cultures by deflned factors. Cell126: 663-676

[43] Tomanck M, Kopecny V, Kanka J (1989) Genome reactivation in developing early embryos: an ultrastructural and autoradiaphic analysis. Anat Embryol180: 309-316

[44] Verma N, Rettenmeier AW, Spanke SS (2011) Recent advances in the use of Sus scrofa (pig) as a model system for proteomic studies. Proteomics11: 776-793

[45] Vignon X, Zhou Q, Renard JP (2002) Chromatin as a regulative architecture od the early developmental functions of mammalian embryos after fertilization or nuclear transfer. Cloning Stem Cells4: 363-377

[46] Wakayama T, Perry ACF, Zuccotti M, Johnson KR, Yanagimachi R (1998) Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature394: 369-374

[47] Walker SC, Shin T, Zaunbrecher GM, Romano JE, Johnson GA, Bazer FW, Piedrahita JA (2002) A highly efflcient method for porcine cloning by nuclear transfer using in vitro-matured oocytes. Cloning Stem Cells4: 105-112

[48] Whitworth K, Prather RS (2010) Somatic cell nuclear transfer efflciency: how can it be improved through nuclear remodeling and reprogramming? Mol Reprod Dev77: 1001-1015

[49] William M, Rideout III, Eggan K, Jaenisch R (2001) Nuclear cloning and epigenetic reprogramming of the genome. Science293: 1093-1098

[50] Wilmut I, Schnieke AE, McWhir J, Kind AJ, Campbell KHS (1997) Viable offspring derived from fetal and adult mammalian cells (vol 385, p 810, 1997). Nature386: 200

[51] Woods GL, White KL, Vanderwall DK, Li GP, Aston KI (2003) A mule cloned from fetal cells by nuclear transfer. Science301: 1063

[52] Yang L, Palmer PC, Tian XC (2005) Expression of imprinted genes is aberrant in deceased newborn cloned calves and relatively normal in surviving adult clones. Mol Reprod Dev71: 431-438

[53] Yu JY, Vodyanik MA, Smuga-Otto K (2007) Induced pluripotent stem cell lines derived from human somatic cells. Science318: 1916-1920

[54] Zhao JG, Hao YH, Prather RS (2010) Histone deacetylase inhibitors improve in vitro and in vivo developmental competence of somatic cell nuclear transfer porcine embryos. Cell Reprogram12: 75-83

[55] Zhou Q, Renard JP, Le Friec G, Brochard V, Beaujean N (2003) Generation of fertile cloned rats by regulating oocyte activation. Science302:1179

RIGHTS & PERMISSIONS

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.