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Tolerance of the Mouse Sperm Nuclei to Freeze-Drying Depends on Their Disulfide Status¹

Institute for Biogenesis Research,³ Department of Anatomy and Reproductive Biology, John A. Burns School of Medicine, University of Hawaii, Honolulu, Hawaii 96822 Division of Reproductive Engineering,⁴ Center for Animal Resources and Development, Kumamoto University, Kumamoto 860-0811, Japan Division of Reproductive Biology,⁵ Department of Obstetrics and Gynecology, University of California, Davis, California 95616

Mouse spermatozoa from the caudae epididymides could be freeze-dried without losing their ability to support normal development. Immature spermatozoa from the testes, in contrast, were damaged by freeze-drying. However, immature spermatozoa became resistant to freeze-drying after their treatment with diamide, which oxidizes free -SH groups. Conversely, epididymal spermatozoa were damaged by freeze-drying if first treated with dithiothreitol (DTT), which reduces -SS- bonds. The potential for freeze-drying damage seems likely to relate to the -SS- status of sperm proteins, in particular its protamines.

¹ This study was conducted as part of the National Cooperative Program on Mouse Sperm Cryopreservation sponsored by the National Institute of Child Health and Human Development and the National Center for Research Resources (U01HD38205).

² Correspondence: Takehito Kaneko, Institute for Biogenesis Research, Department of Anatomy and Reproductive Biology, John A. Burns School of Medicine, University of Hawaii, 1960 East-West Road, Honolulu, HI 96822. FAX: 808 956 7316; takehito{at}hawaii.edu

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