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Enhancement of histone acetylation by trichostatin A during in vitro fertilization of bovine oocytes affects cell number of the inner cell mass of the resulting blastocysts

  • Shuntaro Ikeda (a1), Atsuhiro Tatemizo (a1), Daisaku Iwamoto (a1), Shunji Taniguchi (a2), Yoichiro Hoshino (a3), Tomoko Amano (a1), Kazuya Matsumoto (a1), Yoshihiko Hosoi (a1), Akira Iritani (a1) and Kazuhiro Saeki (a4) (a1)...


Histone acetylation is one of the major mechanisms of epigenetic reprogramming of gamete genomes after fertilization to establish a totipotent state for normal development. In the present study, the effects of trichostatin A (TSA), an inhibitor of histone deacetylase, during in vitro fertilization (IVF) of bovine oocytes on subsequent embryonic development were investigated. Cumulus-enclosed oocytes obtained from slaughterhouse bovine ovaries were matured in vitro and subjected to IVF in a defined medium supplemented with 0 (control), 5, 50, and 500 nM TSA for 18 h. After IVF, presumptive zygotes were cultured in modified synthetic oviductal fluid (mSOF) medium until 168 h postinsemination (hpi). Some oocytes were immunostained using antibody specific for histone H4-acetylated lysine 5 at 10 hpi. Cleavage, blastocyst development and cell number of inner cell mass (ICM) and trophectoderm (TE) of blastocysts were assessed. TSA treatment enhanced histone acetylation that was prominent in decondensed sperm nuclei. TSA did not affect the postfertilization cleavage, blastocyst rates, and TE cell number. However, it significantly enhanced ICM cell number (p < 0.05). These results indicate that TSA treatment during IVF of bovine oocytes does not affect blastocyst development but alters the cell number of ICM, suggesting that overriding epigenetic modification of the genome during fertilization has a carryover effect on cell proliferation and differentiation in preimplantation embryos. Thus, further environmental quality controls in assisted reproductive technologies are needed in terms of factors which affect chromatin remodelling.


Corresponding author

All correspondence to: Kazuhiro Saeki. Department of Genetic Engineering, Kinki University, Wakayama, 6496493, Japan. Tel: +81 736 77 3888. Fax: +81 736 77 4754. e-mail:


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Adenot, P.G., Mercier, Y., Renard, J.P. & Thompson, E.M. (1997). Differential H4 acetylation of paternal and maternal chromatin precedes DNA replication and differential transcriptional activity in pronuclei of 1-cell mouse embryos. Development 124, 4615–25.
Bird, A.P. & Wolffe, A.P. (1999). Methylation-induced repression-belts, braces, and chromatin. Cell 99, 451–4.
Brackett, B.G. & Oliphant, G. (1975). Capacitation of rabbit spermatozoa in vitro. Biol. Reprod. 12, 260–74.
Brison, D.R. & Schultz, R.M. (1997). Apoptosis during mouse blastocyst formation: evidence for a role for survival factors including transforming growth factor alpha. Biol. Reprod. 56, 1088–96.
Cervoni, N. & Szyf, M. (2001). Demethylase activity is directed by histone acetylation. J. Biol. Chem. 276, 40778–87.
Ekwall, K., Olsson, T., Turner, B.M., Cranston, G. & Allshire, R.C. (1997). Transient inhibition of histone deacetylation alters the structural and functional imprint at fission yeast centromeres. Cell 91, 1021–32.
Gioia, L., Barboni, B., Turriani, M., Capacchietti, G., Pistilli, M.G., Berardinelli, P. & Mattioli, M. (2005) The capability of reprogramming the male chromatin after fertilization is dependent on the quality of oocyte maturation. Reproduction 130, 2939.
Holm, S. (1979). A simple sequentially rejective multiple test procedure. Scand. J. Statis. 6, 6570.
Jenuwein, T. & Allis, C.D. (2001). Translating the histone code. Science 293, 1074–80.
Kang, Y.K., Park, J.S., Koo, D.B., Choi, Y.H., Kim, S.U., Lee, K.K. & Han, Y.M. (2002). Limited demethylation leaves mosaic-type methylation states in cloned bovine pre-implantation embryos. EMBO J. 21, 1092–100.
Kasamatsu, A., Saeki, K., Tamari, T., Iwamoto, D., Tatemizo, A., Matsumoto, K., Hosoi, Y. & Iritani, A. (2007). Timing and uniformity of embryonic gene activation affect subsequent pre-implantation development of cloned bovine embryos. J. Reprod. Dev. 53, 623–9.
Kishigami, S., Thuan, N.V., Hikichi, T., Ohta, H., Wakayama, S., Mizutani, E. & Wakayama, T. (2006a). Epigenetic abnormalities of the mouse paternal zygotic genome associated with microinsemination of round spermatids. Dev. Biol. 289, 195205.
Kishigami, S., Mizutani, E., Ohta, H., Hikichi, T., Thuan, N.V., Wakayama, S., Bui, H.T. & Wakayama, T. (2006b). Significant improvement of mouse cloning technique by treatment with trichostatin A after somatic nuclear transfer. Biochem. Biophys. Res. Commun. 340, 183–9.
Koo, D.B., Kang, Y.K., Choi, Y.H., Park, J.S., Kim, H.N., Oh, K.B., Son, D.S., Park, H., Lee, K.K. & Han, Y.M. (2002). Aberrant allocations of inner cell mass and trophectoderm cells in bovine nuclear transfer blastocysts. Biol. Reprod. 67, 487–92.
Maalouf, W.E., Alberio, R. & Campbell, K.H. (2008). Differential acetylation of histone H4 lysine during development of in vitro fertilized, cloned and parthenogenetically activated bovine embryos. Epigenetics 3, 199209.
McGraw, S., Robert, C., Massicotte, L. & Sirard, M. A. (2003). Quantification of histone acetyltransferase and histone deacetylase transcripts during early bovine embryo development. Biol. Reprod. 68, 383–9.
McLay, D.W. & Clarke, H.J. (2003). Remodelling the paternal chromatin at fertilization in mammals. Reproduction 125, 625–33.
Mizzen, C.A. & Allis, C.D. (1998). Linking histone acetylation to transcriptional regulation. Cell. Mol. Life Sci. 54, 620.
Perreault, S.D. (1992). Chromatin remodeling in mammalian zygotes. Mutat. Res. 296, 4355.
Rybouchkin, A., Kato, Y. & Tsunoda, Y. (2006). Role of histone acetylation in reprogramming of somatic nuclei following nuclear transfer. Biol. Reprod. 74, 1083–9.
Saeki, K., Hoshi, M., Leibfried-Rutledge, M.L. & First, N.L. (1990). In vitro fertilization and development of bovine oocytes matured with commercially available follicle stimulating hormone. Theriogenology 34, 1035–9.
Saeki, K., Kato, H., Hosoi, Y., Miyake, M., Utsumi, K. & Iritani, A. (1991). Early morphological events of in vitro fertilized bovine oocytes with frozen–thawed spermatozoa. Theriogenology 35, 1051–8.
Spinaci, M., Seren, E. & Mattioli, M. (2004). Maternal chromatin remodeling during maturation and after fertilization in mouse oocytes. Mol. Reprod. Dev. 69, 215–21.
Takahashi, Y. & First, N.L. (1992). In vitro development of bovine one-cell embryos: Influence of glucose, lactate, pyruvate, amino acids and vitamins. Theriogenology 37, 963–78.
Tarin, J.J., Perez-Albala, S., Gomez-Piquer, V., Hermenegildo, C. & Cano, A. (2002). Stage of the estrous cycle at the time of pregnant mare's serum gonadotropin injection affects pre-implantation embryo development in vitro in the mouse. Mol. Reprod. Dev. 62, 312–9.
Thouas, G.A., Korfiatis, N.A., French, A.J., Jones, G.M. & Trounson, A.O. (2001). Simplified technique for differential staining of inner cell mass and trophectoderm cells of mouse and bovine blastocysts. Reprod. Biomed. Online 3, 25–9.
Torres-Padilla, M.E., Parfitt, D.E., Kouzarides, T. & Zernicka-Goetz, M. (2007). Histone arginine methylation regulates pluripotency in the early mouse embryo. Nature 445, 214–8.
Turner, B.M. (1998). Histone acetylation as an epigenetic determinant of long-term transcriptional competence. Cell. Mol. Life Sci. 54, 2131.
Turner, B.M. (2000). Histone acetylation and an epigenetic code. Bioessays 22, 836–45.
Turner, B.M. (2002). Cellular memory and the histone code. Cell 111, 285–91.
Ward, W.S. & Coffey, D.S. (1991). DNA packaging and organization in mammalian spermatozoa: comparison with somatic cells. Biol. Reprod. 44, 569–74.
Ward, W.S. & Zalensky, A.O. (1996). The unique, complex organization of the transcriptionally silent sperm chromatin. Crit. Rev. Eukaryot. Gene Expr. 6, 139–47.
Wee, G., Koo, D.B., Song, B.S., Kim, J.S., Kang, M.J., Moon, S.J., Kang, Y.K., Lee, K.K. & Han, Y.M. (2006). Inheritable histone H4 acetylation of somatic chromatins in cloned embryos. J. Biol. Chem. 281, 6048–57.



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