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Decrease of Fertilizing Ability of Mouse Spermatozoa after Freezing and Thawing Is Related to Cellular Injury¹

Kyudo Company Limited,³ Kumamoto 861-0104, Japan Center for Animal Resources and Development,⁴ Kumamoto University, Kumamoto 860-0811, Japan Department of Materials and Life Science,⁵ Graduate School of Science and Technology, Kumamoto University, Kumamoto 860-8555, Japan Department of Clinical Chemistry and Informatics,⁶ Graduate School of Medical and Pharmaceutical Sciences, Kumamoto University, Kumamoto 860-8555, Japan

In general, the fertilizing ability of cryopreserved mouse spermatozoa is less than that of fresh spermatozoa. This ability is especially low in C57BL/6, the main strain used for the production of transgenic mice. To solve this problem, the relationship between cell damage and fertilizing ability in cryopreserved mouse spermatozoa was examined in this study. Sperm motility analysis revealed no significant difference among the motilities of cryopreserved C57BL/6J, BALB/cA, and DBA/2N sperm (67.6%, 43.4%, and 60.0%, respectively) after thawing. However, the results of in vitro fertilization (IVF), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) showed a strong correlation between the frequency of aberrant spermatozoa (FAS) and fertilization rates (FR; C57BL/6J: FAS, 83.7%; FR, 17.0%; BALB/cA: FAS, 67.2%; FR, 24.2%; and DBA/2N: FAS, 10.2%; FR, 93.6%), and damage to spermatozoa was localized particularly in the acrosome of the head and mitochondria.

¹ Supported by a Grant-in-Aid for Scientific Research, 11177101-04, from the Japan Society for the Promotion of Science.

² Correspondence: Naomi Nakagata, 2-2-1 Honjo, Kumamoto 860-0811, Japan. FAX: +81 96 373 6560; nakagata{at}gpo.kumamoto-u.ac.jp

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