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Freeze-Dried Sperm Fertilization Leads to Full-Term Development in Rabbits¹

Department of Animal Science/Center for Regenerative Biology,⁵ University of Connecticut, Storrs, Connecticut 06269 Institute for Biogenesis Research,⁶ University of Hawaii Medical School, Honolulu, Hawaii 96822 Faculty of Agriculture and Life Sciences,⁷ Hirosaki University, Hirosaki 036-8561, Japan

To date, the laboratory mouse is the only mammal in which freeze-dried spermatozoa have been shown to support full-term development after microinjection into oocytes. Because spermatozoa in mice, unlike in most other mammals, do not contribute centrosomes to zygotes, it is still unknown whether freeze-dried spermatozoa in other mammals are fertile. Rabbit sperm was selected as a model because of its similarity to human sperm (considering the centrosome inheritance pattern). Freeze- drying induces rabbit spermatozoa to undergo dramatic changes, such as immobilization, membrane breaking, and tail fragmentation. Even when considered to be "dead" in the conventional sense, rabbit spermatozoa freeze-dried and stored at ambient temperature for more than 2 yr still have capability comparable to that of fresh spermatozoa to support preimplantation development after injection into oocytes followed by activation. A rabbit kit derived from a freeze-dried spermatozoon was born after transferring 230 sperm-injected oocytes into eight recipients. The results suggest that freeze-drying could be applied to preserve the spermatozoa from most other species, including human. The present study also raises the question of whether rabbit sperm centrosomes survive freeze-drying or are not essential for embryonic development.

¹ Supported by University of Connecticut Research Foundation (X.C.T. and X.Y.), the Harold Castle Foundation (H.K. and R.Y.), and the Kosasa Family Foundation (H.K. and R.Y.).

² Correspondence. FAX: 860 486 0534; jyang{at}canr.uconn.edu

³ Current address: Carnegie Institution of Washington, Department of Embryology, Baltimore, Maryland 21210

⁴ Current address: Department of Biological Sciences, Asahikawa Medical College, Asahikawa, Hokkaido, Japan

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