Science China Life Sciences

, Volume 55, Issue 4, pp 349–357 | Cite as

Epigenetic reprogramming of embryos derived from sperm frozen at −20°C

  • ShiBin Chao
  • JianChun Li
  • XuanJin Jin
  • HaiXun Tang
  • GongXian Wang
  • GuoLan Gao
Open Access
Research Paper

Abstract

Cryopreservation of spermatozoa is a strategy that has been used to conserve the sperm of animal species and animal strains that are valuable for biomedical research. A simple method for preserving spermatozoa after application of intracytoplasmic sperm injection (ICSI) is much needed. It has been shown previously that spermatozoa frozen at −20°C can activate oocytes and support full-term embryo development. However, epigenetic reprogramming could be affected by the environment and by the in vitro manipulation of gametes. Here, we investigated embryo epigenetic reprogramming including DNA methylation and histone modification, in embryos derived from sperm preserved at −20°C without cryoprotectants. The results showed that although both fertilization and embryo developmental competence were decreased, the dynamic epigenetic reprogramming of embryos derived from frozen sperm was similar to the reprogramming of embryos derived from fresh sperm. The results reported in this study indicate that sperm frozen without cryoprotectant is epigenetically safe for ICSI.

Keywords

spermatozoa freezing ICSI epigenetic histone modification DNA methylation 

References

  1. 1.
    Polge C. Smith A U, Parkes A S. Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature, 1949, 164: 666PubMedCrossRefGoogle Scholar
  2. 2.
    Johnson L A, Aalbers J G, Willems C M, et al. Use of spermatozoa for artificial insemination. I. Fertilizing capacity of fresh and frozen spermatozoa in sows on 36 farms. J Anim Sci, 1981, 52: 1130–1136PubMedGoogle Scholar
  3. 3.
    Gabriel Sanchez-Partida L, Maginnis G, Dominko T, et al. Live rhesus offspring by artificial insemination using fresh sperm and cryopreserved sperm. Biol Reprod, 2000, 63: 1092–1097PubMedCrossRefGoogle Scholar
  4. 4.
    Kelleher S, Wishart S M, Liu P Y, et al. Long-term outcomes of elective human sperm cryostorage. Hum Reprod, 2001, 16: 2632–2639PubMedCrossRefGoogle Scholar
  5. 5.
    Garcia M A, Graham E F. Development of a buffer system for dialysis of bovine spermatozoa before freezing. II. Effect of sugars and sugar alcohols on posthaw motility. Theriogenology, 1989, 31: 1029–1037PubMedCrossRefGoogle Scholar
  6. 6.
    Tada N, Sato M, Yamanoi J, et al. Cryopreservation of mouse spermatozoa in the presence of raffinose and glycerol. J Reprod Fertil, 1990, 89: 511–516PubMedCrossRefGoogle Scholar
  7. 7.
    Kimura Y, Yanagimachi R. Mouse oocytes injected with testicular spermatozoa or round spermatids can develop into normal offspring. Development, 1995, 121: 2397–2405PubMedGoogle Scholar
  8. 8.
    Yanagimachi R. Intracytoplasmic injection of spermatozoa and spermatogenic cells: its biology and applications in humans and animals. Reprod Biomed Online, 2005, 10: 247–288PubMedCrossRefGoogle Scholar
  9. 9.
    Yazawa H, Yanagida K, Hayashi S, et al. The oocyte activation and Ca2+ oscillation-inducing abilities of mouse and human dead (sonicated) spermatozoa. Zygote, 2009, 17: 175–184PubMedCrossRefGoogle Scholar
  10. 10.
    Yan W, Morozumi K, Zhang J, et al. Birth of mice after intracytoplasmic injection of single purified sperm nuclei and detection of messenger RNAs and microRNAs in the sperm nuclei. Biol Reprod, 2008, 78: 896–902PubMedCrossRefGoogle Scholar
  11. 11.
    Wakayama T, Whittingham D G, Yanagimachi R. Production of normal offspring from mouse oocytes injected with spermatozoa cryopreserved with or without cryoprotection. J Reprod Fertil, 1998, 112: 11–17PubMedCrossRefGoogle Scholar
  12. 12.
    Wakayama T, Yanagimachi R. Development of normal mice from oocytes injected with freeze-dried spermatozoa. Nat Biotechnol, 1998, 16: 639–641PubMedCrossRefGoogle Scholar
  13. 13.
    Kusakabe H, Yanagimachi R, Kamiguchi Y. Mouse and human spermatozoa can be freeze-dried without damaging their chromosomes. Hum Reprod, 2008, 23: 233–239PubMedCrossRefGoogle Scholar
  14. 14.
    Keskintepe L, Pacholczyk G, Machnicka A, et al. Bovine blastocyst development from oocytes injected with freeze-dried spermatozoa. Biol Reprod, 2002, 67: 409–415PubMedCrossRefGoogle Scholar
  15. 15.
    Kwon I K, Park K E, Niwa K. Activation, pronuclear formation, and development in vitro of pig oocytes following intracytoplasmic injection of freeze-dried spermatozoa. Biol Reprod, 2004, 71: 1430–1436PubMedCrossRefGoogle Scholar
  16. 16.
    Sanchez-Partida L G, Simerly C R, Ramalho-Santos J. Freeze-dried primate sperm retains early reproductive potential after intracytoplasmic sperm injection. Fertil Steril, 2008, 89: 742–745PubMedCrossRefGoogle Scholar
  17. 17.
    Li M W, Biggers J D, Elmoazzen H Y, et al. Long-term storage of mouse spermatozoa after evaporative drying. Reproduction, 2007, 133: 919–929PubMedCrossRefGoogle Scholar
  18. 18.
    McGinnis L K, Zhu L, Lawitts J A, et al. Mouse sperm desiccated and stored in trehalose medium without freezing. Biol Reprod, 2005, 73: 627–633PubMedCrossRefGoogle Scholar
  19. 19.
    Li C, Mizutani E, Ono T, et al. Intracytoplasmic sperm injection with mouse spermatozoa preserved without freezing for six months can lead to full-term development. Biol Reprod, 2011, 85: 1183–1190PubMedCrossRefGoogle Scholar
  20. 20.
    Ogonuki N, Mochida K, Miki H, et al. Spermatozoa and spermatids retrieved from frozen reproductive organs or frozen whole bodies of male mice can produce normal offspring. Proc Natl Acad Sci USA, 2006, 103: 13098–13103PubMedPubMedCentralCrossRefGoogle Scholar
  21. 21.
    Fleming T P, Kwong W Y, Porter R, et al. The embryo and its future. Biol Reprod, 2004, 71: 1046–1054PubMedCrossRefGoogle Scholar
  22. 22.
    Feng S, Jacobsen S E, Reik W. Epigenetic reprogramming in plant and animal development. Science, 2010, 330: 622–627PubMedPubMedCentralCrossRefGoogle Scholar
  23. 23.
    Ballestar, E. An introduction to epigenetics. Adv Exp Med Biol, 2011, 711: 1–11PubMedCrossRefGoogle Scholar
  24. 24.
    Kimura Y, Yanagimachi R. Intracytoplasmic sperm injection in the mouse. Biol Reprod, 1995, 52: 709–720PubMedCrossRefGoogle Scholar
  25. 25.
    Yoshida N, Perry A C F. Piezo-actuated mouse intracytoplasmic sperm injection (ICSI). Nat Protoc, 2007, 2: 296–304PubMedCrossRefGoogle Scholar
  26. 26.
    Kamiguchi Y, Mikamo K. An improved, efficient method for analyzing human sperm chromosomes using zona-free hamster ova. Am J Hum Genet, 1986, 38: 724–740PubMedPubMedCentralGoogle Scholar
  27. 27.
    Akiyama T, Nagata M, Aoki F. Inadequate histone deacetylation during oocyte meiosis causes aneuploidy and embryo death in mice. Proc Natl Acad Sci USA, 2006, 103: 7339–7344PubMedPubMedCentralCrossRefGoogle Scholar
  28. 28.
    Kusakabe H, Szczygiel M A, Whittingham D G, et al. Maintenance of genetic integrity in frozen and freeze-dried mouse spermatozoa. Proc Natl Acad Sci USA, 2001, 98: 13501–13506PubMedPubMedCentralCrossRefGoogle Scholar
  29. 29.
    Kaneko T, Whittingham D G, Yanagimachi R. Effect of pH value of freeze-drying solution on the chromosome integrity and developmental ability of mouse spermatozoa. Biol Reprod, 2003, 68: 136–139PubMedCrossRefGoogle Scholar
  30. 30.
    Kim J M, Ogura A, Nagata M, et al. Analysis of the mechanism for chromatin remodeling in embryos reconstructed by somatic nuclear transfer. Biol Reprod, 2002, 67: 760–766PubMedCrossRefGoogle Scholar
  31. 31.
    Lacham-Kaplan O, Shaw J, Sanchez-Partida L G, et al. Oocyte activation after intracytoplasmic injection with sperm frozen without cryoprotectants results in live offspring from inbred and hybrid mouse strains. Biol Reprod, 2003, 69: 1683–1689PubMedCrossRefGoogle Scholar
  32. 32.
    Santos F, Hendrich B, Reik W, et al. Dynamic reprogramming of DNA methylation in the early mouse embryo. Dev Biol, 2002, 241: 172–182PubMedCrossRefGoogle Scholar
  33. 33.
    Mayer W, Niveleau A, Walter J, et al. Demethylation of the zygotic paternal genome. Nature, 2000, 403: 501–502PubMedCrossRefGoogle Scholar
  34. 34.
    Dean W, Santos F, Stojkovic M, et al. Conservation of methylation reprogramming in mammalian development: aberrant reprogramming in cloned embryos. Proc Natl Acad Sci USA, 2001, 98: 13734–13738PubMedPubMedCentralCrossRefGoogle Scholar
  35. 35.
    Metivier R, Gallais R, Tiffoche C, et al. Cyclical DNA methylation of a transcriptionally active promoter. Nature, 2008, 452: 45–50PubMedCrossRefGoogle Scholar
  36. 36.
    Zaitseva I, Zaitsev S, Alenina N, et al. Dynamics of DNA-demethylation in early mouse and rat embryos developed in vivo and in vitro. Mol Reprod Dev, 2007, 74: 1255–1261PubMedCrossRefGoogle Scholar
  37. 37.
    Lepikhov K, Walter J. Differential dynamics of histone H3 methylation at positions K4 and K9 in the mouse zygote. BMC Dev Biol, 2004, 4: 12PubMedPubMedCentralCrossRefGoogle Scholar
  38. 38.
    Lachner M, Jenuwein T. The many faces of histone lysine methylation. Curr Opin Cell Biol, 2002, 14: 286–298PubMedCrossRefGoogle Scholar
  39. 39.
    Strauss G, Schurtenberger P, Hauser H. The interaction of saccharides with lipid bilayer vesicles: stabilization during freeze-thawing and freeze-drying. Biochim Biophys Acta, 1986, 858: 169–180PubMedCrossRefGoogle Scholar
  40. 40.
    Yan W, Morozumi K, Zhang J, et al. Birth of mice after intracytoplasmic injection of single purified sperm nuclei and detection of messenger RNAs and MicroRNAs in the sperm nuclei. Biol Reprod, 2008, 78: 896–902PubMedCrossRefGoogle Scholar
  41. 41.
    Miller D, Brinkworth M, Iles D. Paternal DNA packaging in spermatozoa: more than the sum of its parts? DNA, histones, protamines and epigenetics. Reproduction, 2010, 139: 287–301PubMedCrossRefGoogle Scholar
  42. 42.
    Yildiz C, Ottaviani P, Law N, et al. Effects of cryopreservation on sperm quality, nuclear DNA integrity, in vitro fertilization, and in vitro embryo development in the mouse. Reproduction, 2007, 133: 585–595PubMedCrossRefGoogle Scholar
  43. 43.
    Feng S, Jacobsen S E, Reik W. Epigenetic reprogramming in plant and animal development. Science, 2010, 330: 622–627PubMedPubMedCentralCrossRefGoogle Scholar
  44. 44.
    Vassena R, Dee Schramm R, Latham K E. Species-dependent expression patterns of DNA methyltransferase genes in mammalian oocytes and preimplantation embryos. Mol Reprod Dev, 2005, 72: 430–436PubMedCrossRefGoogle Scholar
  45. 45.
    Feil R. Epigenetic asymmetry in the zygote and mammalian development. Int J Dev Biol, 2009, 53: 191–201PubMedCrossRefGoogle Scholar
  46. 46.
    Klose R J, Zhang Y. Regulation of histone methylation by demethylimination and demethylation. Nat Rev Mol Cell Biol, 2007, 8: 307–318PubMedCrossRefGoogle Scholar
  47. 47.
    Kouzarides T. Chromatin modifications and their function. Cell, 2007, 128: 693–705PubMedCrossRefGoogle Scholar
  48. 48.
    Erhardt S, Su I H, Schneider R, et al. Consequences of the depletion of zygotic and embryonic enhancer of zeste 2 during preimplantation mouse development. Development, 2003, 130: 4235–4248PubMedCrossRefGoogle Scholar
  49. 49.
    Linggi B E, Brandt S J, Sun Z W, et al. Translating the histone code into leukemia. J Cell Biochem, 2005, 96: 938–950PubMedCrossRefGoogle Scholar
  50. 50.
    Kishigami S, Van Thuan N, Hikichi T, et al. Epigenetic abnormalities of the mouse paternal zygotic genome associated with microinsemination of round spermatids. Dev Biol, 2006, 289: 195–205PubMedCrossRefGoogle Scholar

Copyright information

© The Author(s) 2012

Authors and Affiliations

  • ShiBin Chao
    • 1
    • 2
  • JianChun Li
    • 1
  • XuanJin Jin
    • 1
  • HaiXun Tang
    • 1
  • GongXian Wang
    • 1
  • GuoLan Gao
    • 2
  1. 1.Center of Reproductive Medicinethe First Affiliated Hospital of Nanchang UniversityNanchangChina
  2. 2.Department of Obstetrics and GynecologyAviation General Hospital of ChinaBeijingChina

Personalised recommendations