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Sperm Aster Formation and Pronuclear Decondensation During Rabbit Fertilization and Development of a Functional Assay for Human Sperm¹

^a Department of Reproductive Sciences, Oregon Regional Primate Research Center (Oregon Health Sciences University), Beaverton, Oregon 97006 ^b Department of Cell-Developmental Biology and ^c Department of Obstetrics and Gynecology, Oregon Health Sciences University, Portland, Oregon 97201

Microtubule organization and chromatin configurations in rabbit eggs after in vivo rabbit fertilization and after intracytoplasmic injection with human sperm were characterized. In unfertilized eggs, an anastral barrel-shaped meiotic spindle, oriented radially to the cortex, was observed. After rabbit sperm incorporation, microtubules were organized into a radial aster from the sperm head, and cytoplasmic microtubules were organized around the male and female pronuclei. The microtubules extending from the decondensed sperm head participated in pronuclear migration, and organization around the female pronucleus may also be important for pronuclear centration. Support for these observations was found in parthenogenetically activated eggs, in which microtubule arrays were organized around the single female pronucleus that formed after artificial activation. These observations support a biparental centrosomal contribution during rabbit fertilization as opposed to a strictly paternal inheritance pattern suggested from previous studies. In rabbit eggs that received injected human donor sperm, an astral array of microtubules radiated from the sperm neck and enlarged as the sperm head underwent pronuclear decondensation. -Tubulin was observed in the center of the sperm aster. We conclude that the rabbit egg exhibits a blended centrosomal contribution necessary for completion of fertilization and that the rabbit egg may be a novel animal model for assessing centrosomal function in human sperm and spermatogenic cells following intracytoplasmic injection.

¹ This investigation was supported by research grants from the National Institutes of Health (NICHD, NCRR), and by NICHD/NIH through cooperative agreement U54 HD18185-16 as part of the Specialized Cooperative Centers Program in Reproduction Research. The Oregon Regional Primate Research Center (ORPRC) is sponsored as an NIH/NCRR Regional Primate Research Center.

² Correspondence: Gerald Schatten, 505 N.W. 185th Avenue, Beaverton, OR 97006. FAX: 503 614 3725; schatten{at}ohsu.edu

³ Permanent address: Yukihiro Terada, Department of Obstetrics and Gynecology, Tohoku University School of Medicine, Sendai, Japan.

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