Hostname: page-component-76fb5796d-2lccl Total loading time: 0 Render date: 2024-04-26T02:42:53.030Z Has data issue: false hasContentIssue false
  • English
  • Français

Preservation of sperm within the mouse cauda epididymidis in salt or sugars at room temperature

Published online by Cambridge University Press:  29 January 2010

Tetsuo Ono ,
Eiji Mizutani ,
Chong Li  and
Teruhiko Wakayama
Tetsuo Ono
Affiliation:
Laboratory for Genomic Reprogramming, RIKEN Center for Developmental Biology, Kobe 650–0047, Japan. Department of Medical Science, Graduate School of Medicine, Kyoto University, Kyoto 606–8501, Japan.
Eiji Mizutani
Affiliation:
Laboratory for Genomic Reprogramming, RIKEN Center for Developmental Biology, Kobe 650–0047, Japan.
Chong Li
Affiliation:
Laboratory for Genomic Reprogramming, RIKEN Center for Developmental Biology, Kobe 650–0047, Japan. Department of Bioscience, Graduate School of Science and Technology, Kwansei Gakuin University, Sanda 662–8501, Japan.
Teruhiko Wakayama*
Affiliation:
Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN, 2–2-3 Minatojima-minamimachi, Chuo-ku, Kobe 650–0047, Japan. Laboratory for Genomic Reprogramming, RIKEN Center for Developmental Biology, Kobe 650–0047, Japan. Department of Medical Science, Graduate School of Medicine, Kyoto University, Kyoto 606–8501, Japan. Department of Bioscience, Graduate School of Science and Technology, Kwansei Gakuin University, Sanda 662–8501, Japan.
*
All correspondence to: Teruhiko Wakayama. Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN, 2–2-3 Minatojima-minamimachi, Chuo-ku, Kobe 650–0047, Japan. Tel: +81 78 306 3049. Fax: +81 78 306 3095. e-mail: teru@cdb.riken.jp
Get access Rights & Permissions [Opens in a new window]

Summary

The development of preservation techniques for male gametes at room temperature might allow us to store them in a simple and cost-effective manner. In this study, we studied the use of pure salt or sugar to preserve the whole cauda epididymidis, because it is known that food can be preserved in this way at room temperature for long periods. Mouse epididymides were placed directly in powdered salt (NaCl) or sugars (glucose or raffinose) for 1 day to 1 year at room temperature. Spermatozoa were recovered from the preserved organs after being rehydrated with medium and then isolated sperm heads were microinjected into fresh oocytes. Importantly, the oocyte activation capacity of spermatozoa was maintained after epididymal storage in NaCl for 1 year, whereas most untreated spermatozoa failed to activate oocytes within 1 month of storage. Pronuclear morphology, the rate of extrusion of a second polar body and the methylation status of histone H3 lysine 9 (H3K9me3) in those zygotes were similar to those of zygotes fertilized with fresh spermatozoa. However, the developmental ability of the zygotes decreased within 1 day of sperm storage. This effect led to nuclear fragmentation at the 2-cell embryo stage, irrespective of the storage method used. Thus, although the preserved sperm failed to allow embryo development, their oocyte activation factors were maintained by salt storage of the epididymis for up to 1 year at room temperature.

Keywords

ICSIPreservationSaltSpermSugar
Type
Research Article
Information
Zygote , Volume 18 , Issue 3 , August 2010 , pp. 245 - 256
Copyright
Copyright © Cambridge University Press 2010

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Abe, Y., Lee, D.S., Sano, H., Akiyama, K., Yanagimoto-Ueta, Y., Asano, T., Suwa, Y. & Suzuki, H. (2008). Artificial insemination with canine spermatozoa frozen in a skim milk/glucose-based extender. J. Reprod. Dev. 54, 290–4.CrossRefGoogle Scholar
Acott, T.S. & Carr, D.W. (1984). Inhibition of bovine spermatozoa by caudal epididymal fluid. II. Interaction of pH and a quiescence factor. Biol. Reprod. 30, 926–35.CrossRefGoogle Scholar
Chian, R.C., Niwa, K. & Okuda, K. (1991). In vitro penetration of zona pellucida of salt-stored bovine oocytes before and after maturation by frozen–thawed spermatozoa. Theriogenology 36, 209–19.CrossRefGoogle ScholarPubMed
Cox, L.J., Larman, M.G., Saunders, C.M., Hashimoto, K., Swann, K. & Lai, F.A. (2002). Sperm phospholipase Czeta from humans and cynomolgus monkeys triggers Ca2+ oscillations, activation and development of mouse oocytes. Reproduction 124, 611–23.CrossRefGoogle ScholarPubMed
De Pauw, I.M., Van Soom, A., Mintiens, K., Verberckmoes, S. & de Kruif, A. (2003). In vitro survival of bovine spermatozoa stored at room temperature under epididymal conditions. Theriogenology 59, 1093–107.CrossRefGoogle ScholarPubMed
Fan, Z.Q., Li, X.W., Liu, Y., Meng, Q.G., Wang, Y.P., Hou, Y.P., Zhou, G.B. & Zhu, S.E. (2008). Piezo-assisted in vitro fertilization of mouse oocytes with spermatozoa retrieved from epididymides stored at 4 °C. J. Reprod. Dev. 54, 107–12.CrossRefGoogle Scholar
Gatti, J.L., Druart, X., Syntin, P., Guerin, Y., Dacheux, J.L. & Dacheux, F. (2000). Biochemical characterization of two ram cauda epididymal maturation-dependent sperm glycoproteins. Biol. Reprod. 62, 950–8.CrossRefGoogle ScholarPubMed
Hishinuma, M., Suzuki, K. & Sekine, J. (2003). Recovery and cryopreservation of sika deer (Cervus nippon) spermatozoa from epididymides stored at 4 °C. Theriogenology 59, 813–20.CrossRefGoogle Scholar
Kaabi, M., Paz, P., Alvarez, M., Anel, E., Boixo, J.C., Rouissi, H., Herraez, P. & Anel, L. (2003). Effect of epididymis handling conditions on the quality of ram spermatozoa recovered post-mortem. Theriogenology 60, 1249–59.CrossRefGoogle ScholarPubMed
Kikuchi, K.K.N., Nagai, T., Noguchi, J., Shimada, A., Takahashi, R., Hirabayashi, M., Shino, M., Ueda, M. & Kaneko, H. (1999). Reproduction in pigs using frozen–thawed spermatozoa from epididymis stored at 4 °C. J. Reprod. Dev. 45, 345–50.CrossRefGoogle Scholar
Kimura, Y. & Yanagimachi, R. (1995). Intracytoplasmic sperm injection in the mouse. Biol. Reprod. 52, 709–720.CrossRefGoogle ScholarPubMed
Kimura, Y., Yanagimachi, R., Kuretake, S., Bortkiewicz, H., Perry, A.C. & Yanagimachi, H. (1998). Analysis of mouse oocyte activation suggests the involvement of sperm perinuclear material. Biol. Reprod. 58, 1407–15.CrossRefGoogle ScholarPubMed
Martin-DeLeon, P.A. (2006). Epididymal SPAM1 and its impact on sperm function. Mol. Cell. Endocrinol. 250, 114–21.CrossRefGoogle ScholarPubMed
Martinez-Pastor, F., Guerra, C., Kaabi, M., Diaz, A.R., Anel, E., Herraez, P., de Paz, P. & Anel, L. (2005). Decay of sperm obtained from epididymes of wild ruminants depending on postmortem time. Theriogenology 63, 24–40.CrossRefGoogle ScholarPubMed
McGinnis, L.K., Zhu, L., Lawitts, J.A., Bhowmick, S., Toner, M. & Biggers, J.D. (2005). Mouse sperm desiccated and stored in trehalose medium without freezing. Biol. Reprod. 73, 627–33.CrossRefGoogle ScholarPubMed
Ohta, H., Sakaide, Y. & Wakayama, T. (2008). Long-term preservation of mouse spermatozoa as frozen testicular sections. J. Reprod. Dev. 54, 295–8.CrossRefGoogle ScholarPubMed
Ono, T., Mizutani, E., Li, C. & Wakayama, T. (2008). Nuclear transfer preserves the nuclear genome of freeze-dried mouse cells. J. Reprod. Dev. 54, 486–91.CrossRefGoogle ScholarPubMed
Saunders, C. M., Larman, M. G., Parrington, J., Cox, L. J., Royse, J., Blayney, L. M., Swann, K. & Lai, F. A. (2002). PLC zeta: a sperm-specific trigger of Ca2+ oscillations in eggs and embryo development. Development 129, 3533–44.CrossRefGoogle ScholarPubMed
Sette, C., Bevilacqua, A., Bianchini, A., Mangia, F., Geremia, R. & Rossi, P. (1997). Parthenogenetic activation of mouse eggs by microinjection of a truncated c-kit tyrosine kinase present in spermatozoa. Development 124, 2267–74.CrossRefGoogle ScholarPubMed
Sette, C., Paronetto, M.P., Barchi, M., Bevilacqua, A., Geremia, R. & Rossi, P. (2002). Tr-kit-induced resumption of the cell cycle in mouse eggs requires activation of a Src-like kinase. EMBO J. 21, 5386–95.CrossRefGoogle ScholarPubMed
Soler, A.J., Perez-Guzman, M.D. & Garde, J.J. (2003). Storage of red deer epididymides for four days at 5°C: effects on sperm motility, viability and morphological integrity. J. Exp. Zoolog. A Comp. Exp. Biol. 295, 188–99.CrossRefGoogle Scholar
Sztein, J.M., Farley, J.S. & Mobraaten, L.E. (2000). In vitro fertilization with cryopreserved inbred mouse sperm. Biol. Reprod. 63, 1774–80.CrossRefGoogle ScholarPubMed
Tateno, H., Wakayama, T., Ward, W.S. & Yanagimachi, R. (1998). Can alcohol retain the reproductive and genetic potential of sperm nuclei? Chromosome analysis of mouse spermatozoa stored in alcohol. Zygote 6, 233–8.CrossRefGoogle Scholar
Van Thuan, N., Wakayama, S., Kishigami, S. & Wakayama, T. (2005). New preservation method for mouse spermatozoa without freezing. Biol. Reprod. 72, 444–50.CrossRefGoogle ScholarPubMed
Wakayama, T. & Yanagimachi, R. (1998). Development of normal mice from oocytes injected with freeze-dried spermatozoa. Nat. Biotechnol. 16, 639–41.CrossRefGoogle ScholarPubMed
Ward, M.A., Kaneko, T., Kusakabe, H., Biggers, J.D., Whittingham, D.G. & Yanagimachi, R. (2003). Long-term preservation of mouse spermatozoa after freeze-drying and freezing without cryoprotection. Biol. Reprod. 69, 2100–8.CrossRefGoogle ScholarPubMed
Wijnker, J.J., Koop, G. & Lipman, L.J. (2006). Antimicrobial properties of salt (NaCl) used for the preservation of natural casings. Food Microbiol. 23, 657–62.CrossRefGoogle ScholarPubMed
Wu, A.T., Sutovsky, P., Manandhar, G., Xu, W., Katayama, M., Day, B.N., Park, K. W., Yi, Y.J., Xi, Y.W., Prather, R.S. & Oko, R. (2007). PAWP, a sperm-specific WW domain-binding protein, promotes meiotic resumption and pronuclear development during fertilization. J. Biol. Chem. 282, 12164–75.CrossRefGoogle ScholarPubMed
Xu, W., Ensrud, K.M. & Hamilton, D.W. (1997). The 26 kD protein recognized on rat cauda epididymal sperm by monoclonal antibody 4E9 has internal peptide sequence that is identical to the secreted form of epididymal protein E. Mol. Reprod. Dev. 46, 377–82.3.0.CO;2-W>CrossRefGoogle Scholar
Yamagata, K., Suetsugu, R. & Wakayama, T. (2009). Long-term, six-dimensional live-cell imaging for the mouse preimplantation embryo that does not affect full-term development. J. Reprod. Dev. 55, 343–50.CrossRefGoogle Scholar
Yanagimachi, R., Lopata, A., Odom, C.B., Bronson, R.A., Mahi, C.A. & Nicolson, G.L. (1979). Retention of biologic characteristics of zona pellucida in highly concentrated salt solution: the use of salt-stored eggs for assessing the fertilizing capacity of spermatozoa. Fertil. Steril. 31, 562–74.CrossRefGoogle ScholarPubMed
Yu, I. & Leibo, S.P. (2002). Recovery of motile, membrane-intact spermatozoa from canine epididymides stored for 8 days at 4 °C. Theriogenology 57, 1179–90.CrossRefGoogle Scholar
Zhang, M., Tang, H., Shen, G., Zhou, B., Wu, Z., Peng, Z., Zhang, J., Yan, J. & Xia, G. (2005). Atrial natriuretic peptide induces an acrosome reaction in giant panda spermatozoa and enhances their penetration of salt-stored porcine oocytes. Theriogenology 64, 1297–308.CrossRefGoogle ScholarPubMed