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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: [email protected]
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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

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