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Defective Histone H3K27 Trimethylation Modification in Embryos Derived from Heated Mouse Sperm

  • Shi-Bin Chao (a1) (a2), Lei Chen (a2), Jian-Chun Li (a1), Xiang-Hong Ou (a2), Xiao-Jin Huang (a1), Shu Wen (a3), Qing-Yuan Sun (a2) and Guo-Lan Gao (a1) (a4)...


The mouse sperm genome is resistant to in vitro heat treatment, and embryos derived from heated sperm can support full-term embryonic development, but the blastocyst rate and implantation rate are lower compared to embryos derived from fresh sperm. In the present study, the patterns of DNA methylation, histone H4K12 (ACH4K12) acetylation, H3K9 trimethylation (H3K9-TriM), and H3K27 trimethylation (H3K27-TriM) in preimplantation embryos derived from 65°C-heated sperm were investigated. Although no evident changes in global DNA methyaltion, histone H4K12 (ACH4K12) acetylation, and H3K9 trimethylation (H3K9-TriM) were found, significantly lower levels of H3K27-TriM, which was thought to be one of the reasons for low efficiency of mouse cloning, were found in the inner cell mass of heated-sperm derived blastocysts. Thus, defective modification of H3K27-TriM might contribute to compromised development of embryos derived from heated sperm.


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Ball, M.J. (1987). Effect of two disinfectant treatments on laboratory analyses. J R Soc Med 80(8), 482484.
Banks, S., King, S.A., Irvine, D.S. & Saunders, P.T. (2005). Impact of a mild scrotal heat stress on DNA integrity in murine spermatozoa. Reproduction 129(4), 505514.
Bernstein, B.E., Mikkelsen, T.S., Xie, X., Kamal, M., Huebert, D.J., Cuff, J., Fry, B., Meissner, A., Wernig, M., Plath, K., Jaenisch, R., Wagschal, A., Feil, R., Schreiber, S.L. & Lander, E.S. (2006). A bivalent chromatin structure marks key developmental genes in embryonic stem cells. Cell 125(2), 315326.
Cao, R., Wang, L., Wang, H., Xia, L., Erdjument-Bromage, H., Tempst, P., Jones, R.S. & Zhang, Y. (2002). Role of histone H3 lysine 27 methylation in Polycomb-group silencing. Science 298(5595), 10391043.
Chambers, I., Colby, D., Robertson, M., Nichols, J., Lee, S., Tweedie, S. & Smith, A. (2003). Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 113(5), 643655.
Chao, S.B., Guo, L., Ou, X.H., Luo, S.M., Wang, Z.B., Schatten, H., Gao, G.L. & Sun, Q.Y. (2012). Heated spermatozoa: Effects on embryonic development and epigenetics. Hum Reprod 27(4), 10161024.
Cozzi, J., Monier-Gavelle, F., Lievre, N., Bomsel, M. & Wolf, J.P. (2001). Mouse offspring after microinjection of heated spermatozoa. Biol Reprod 65(5), 15181521.
Dean, W., Santos, F., Stojkovic, M., Zakhartchenko, V., Walter, J., Wolf, E. & Reik, W. (2001). Conservation of methylation reprogramming in mammalian development: aberrant reprogramming in cloned embryos. Proc Natl Acad Sci USA 98(24), 1373413738.
Erhardt, S., Su, I.H., Schneider, R., Barton, S., Bannister, A.J., Perez-Burgos, L., Jenuwein, T., Kouzarides, T., Tarakhovsky, A. & Surani, M.A. (2003). Consequences of the depletion of zygotic and embryonic enhancer of zeste 2 during preimplantation mouse development. Development 130(18), 42354248.
Feil, R. (2009). Epigenetic asymmetry in the zygote and mammalian development. Int J Dev Biol 53(2-3), 191201.
Feng, S., Jacobsen, S.E. & Reik, W. (2010). Epigenetic reprogramming in plant and animal development. Science 330(6004), 622627.
Fleming, T.P., Kwong, W.Y., Porter, R., Ursell, E., Fesenko, I., Wilkins, A., Miller, D.J., Watkins, A.J. & Eckert, J.J. (2004). The embryo and its future. Biol Reprod 71(4), 10461054.
Jiang, M.X., Zhu, Y., Zhu, Z.Y., Sun, Q.Y. & Chen, D.Y. (2005). Effects of cooling, cryopreservation and heating on sperm proteins, nuclear DNA, and fertilization capability in mouse. Mol Reprod Dev 72(1), 129134.
Jones, P.A. & Takai, D. (2001). The role of DNA methylation in mammalian epigenetics. Science 293(5532), 10681070.
Kaneko, T., Whittingham, D.G. & Yanagimachi, R. (2003). Effect of pH value of freeze-drying solution on the chromosome integrity and developmental ability of mouse spermatozoa. Biol Reprod 68(1), 136139.
Khosla, S., Dean, W., Brown, D., Reik, W. & Feil, R. (2001). Culture of preimplantation mouse embryos affects fetal development and the expression of imprinted genes. Biol Reprod 64(3), 918926.
Kim, J.M., Ogura, A., Nagata, M. & Aoki, F. (2002). Analysis of the mechanism for chromatin remodeling in embryos reconstructed by somatic nuclear transfer. Biol Reprod 67(3), 760766.
Kimura, Y. & Yanagimachi, R. (1995). Intracytoplasmic sperm injection in the mouse. Biol Reprod 52(4), 709720.
Klose, R.J. & Zhang, Y. (2007). Regulation of histone methylation by demethylimination and demethylation. Nat Rev Mol Cell Biol 8(4), 307318.
Kouzarides, T. (2007). Chromatin modifications and their function. Cell 128(4), 693705.
Kwon, I.K., Park, K.E. & Niwa, K. (2004). Activation, pronuclear formation, and development in vitro of pig oocytes following intracytoplasmic injection of freeze-dried spermatozoa. Biol Reprod 71(5), 14301436.
Lachner, M. & Jenuwein, T. (2002). The many faces of histone lysine methylation. Curr Opin Cell Biol 14(3), 286298.
Lepikhov, K. & Walter, J. (2004). Differential dynamics of histone H3 methylation at positions K4 and K9 in the mouse zygote. BMC Dev Biol 4, 12.
Li, C., Mizutani, E., Ono, T. & Wakayama, T. (2009). Production of normal mice from spermatozoa denatured with high alkali treatment before ICSI. Reproduction 137(5), 779792.
Li, E. (2002). Chromatin modification and epigenetic reprogramming in mammalian development. Nat Rev Genet 3(9), 662673.
Mayer, W., Niveleau, A., Walter, J., Fundele, R. & Haaf, T. (2000). Demethylation of the zygotic paternal genome. Nature 403(6769), 501502.
Metivier, R., Gallais, R., Tiffoche, C., Le Peron, C., Jurkowska, R.Z., Carmouche, R.P., Ibberson, D., Barath, P., Demay, F., Reid, G., Benes, V., Jeltsch, A., Gannon, F. & Salbert, G. (2008). Cyclical DNA methylation of a transcriptionally active promoter. Nature 452(7183), 4550.
Miller, D., Brinkworth, M. & Iles, D. (2009). Paternal DNA packaging in spermatozoa: More than the sum of its parts? DNA, histones, protamines and epigenetics. Reproduction 139(2), 287301.
Mitsui, K., Tokuzawa, Y., Itoh, H., Segawa, K., Murakami, M., Takahashi, K., Maruyama, M., Maeda, M. & Yamanaka, S. (2003). The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell 113(5), 631642.
Morgan, H.D., Santos, F., Green, K., Dean, W. & Reik, W. (2005). Epigenetic reprogramming in mammals. Hum Mol Genet 14(1), R47R58.
Paul, C., Murray, A.A., Spears, N. & Saunders, P.T. (2008). A single, mild, transient scrotal heat stress causes DNA damage, subfertility and impairs formation of blastocysts in mice. Reproduction 136(1), 7384.
Santos, F., Hendrich, B., Reik, W. & Dean, W. (2002). Dynamic reprogramming of DNA methylation in the early mouse embryo. Dev Biol 241(1), 172182.
Surani, M.A., Hayashi, K. & Hajkova, P. (2007). Genetic and epigenetic regulators of pluripotency. Cell 128(4), 747762.
Wakayama, T. & Yanagimachi, R. (1998). Development of normal mice from oocytes injected with freeze-dried spermatozoa. Nat Biotechnol 16(7), 639641.
Ward, W.S., Kimura, Y. & Yanagimachi, R. (1999). An intact sperm nuclear matrix may be necessary for the mouse paternal genome to participate in embryonic development. Biol Reprod 60(3), 702706.
Yan, W., Morozumi, K., Zhang, J., Ro, S., Park, C. & Yanagimachi, R. (2008). Birth of mice after intracytoplasmic injection of single purified sperm nuclei and detection of messenger RNAs and MicroRNAs in the sperm nuclei. Biol Reprod 78(5), 896902.
Yanagida, K., Yanagimachi, R., Perreault, S.D. & Kleinfeld, R.G. (1991). Thermostability of sperm nuclei assessed by microinjection into hamster oocytes. Biol Reprod 44(3), 440447.
Yanagimachi, R. (2011). Mammalian sperm acrosome reaction: Where does it begin before fertilization? Biol Reprod 85(1), 45.
Yazawa, H., Yanagida, K., Hayashi, S. & Sato, A. (2009). The oocyte activation and Ca2+ oscillation-inducing abilities of mouse and human dead (sonicated) spermatozoa. Zygote 17(2), 175184.
Yeo, S., Lee, K.K., Han, Y.M. & Kang, Y.K. (2005). Methylation changes of lysine 9 of histone H3 during preimplantation mouse development. Mol Cells 20(3), 423428.
Yoshida, N. & Perry, A.C.F. (2007). Piezo-actuated mouse intracytoplasmic sperm injection (ICSI). Nat Protoc 2(2), 296304.
Zhang, M., Wang, F., Kou, Z., Zhang, Y. & Gao, S. (2009). Defective chromatin structure in somatic cell cloned mouse embryos. J Biol Chem 284(37), 2498124987.



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