Biol Reprod Keystone Symposia Conference on Frontiers in Reproductive Biology & Regulation of Fertility.
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BOR - Papers in Press, published online ahead of print February 11, 2004.
Biol Reprod 2004, 10.1095/biolreprod.103.025957
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Submitted November 26, 2003
Returned for revision December 12, 2003
Accepted February 2, 2004

Reproductive Technology


Freeze-Dried Sperm Fertilization Leads to Term Development in Rabbits

Ji-Long Liu , Hirokazu Kusakabe , Ching-Chien Chang , Hiroyuki Suzuki , David W. Schmidt , Marina Julian , Robert Pfeffer , Charles L. Bormann , X. Cindy Tian , Ryuzo Yanagimachi , and Xiangzhong Yang *

* To whom correspondence should be addressed. E-mail: jyang{at}canr.uconn.edu.

Abstract
To date, the laboratory mouse is the only mammals 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 or not. 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 'dead' in the conventional sense, rabbit spermatozoa freeze-dried and stored at ambient temperature for more than two years still have comparable capability as fresh spermatozoa to support pre-implantation 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 8 recipients. The results suggest that freeze-drying could be applied to preserve the spermatozoa from most other species including human. This study also raises the question of whether rabbit sperm centrosomes survive freeze-drying or are not essential for embryonic development.

Key words: Fertilization • In vitro fertilization • Oocyte development • Sperm


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