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Vol. 10, Issue 9, 2955-2969, September 1999

Biparental Inheritance of -Tubulin during Human Fertilization: Molecular Reconstitution of Functional Zygotic Centrosomes in Inseminated Human Oocytes and in Cell-free Extracts Nucleated by Human Sperm

Calvin Simerly,^* Sara S. Zoran,^* Chris Payne,^* Tanja Dominko,^* Peter Sutovsky,^* Christopher S. Navara,^* Jeffery L. Salisbury, and Gerald Schatten^*

 ^*Departments of Cell-Developmental Biology and Obstetrics-Gynecology, Oregon Health Sciences University, and the Oregon Regional Primate Research Center, Portland, Oregon 97006; and  Department of Biochemistry and Molecular Biology, Mayo Clinic Foundation, Rochester, Minnesota 55905

Human sperm centrosome reconstitution and the parental contributions to the zygotic centrosome are examined in mammalian zygotes and after exposure of spermatozoa to Xenopus laevis cell-free extracts. The presence and inheritance of the conserved centrosomal constituents -tubulin, centrin, and MPM-2 (which detects phosphorylated epitopes) are traced, as is the sperm microtubule-nucleating capability on reconstituted centrosomes. -Tubulin is biparentally inherited in humans (maternal >> than paternal): Western blots detect the presence of paternal -tubulin. Recruitment of maternal -tubulin to the sperm centrosome occurs after sperm incorporation in vivo or exposure to cell-free extract, especially after sperm "priming" induced by disulfide bond reduction. Centrin is found in the proximal sperm centrosomal region, demonstrates expected calcium sensitivity, but appears absent from the zygotic centrosome after sperm incorporation or exposure to extracts. Sperm centrosome phosphorylation is detected after exposure of primed sperm to egg extracts as well as during the early stages of sperm incorporation after fertilization. Finally, centrosome reconstitution in cell-free extracts permits sperm aster microtubule assembly in vitro. Collectively, these results support a model of a blended zygotic centrosome composed of maternal constituents attracted to an introduced paternal template after insemination.

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   Corresponding author. E-mail address: schatten{at}ohsu.edu.

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Molecular Biology of the Cell
Vol. 10, 2955-2969, September 1999
Copyright © 1999 by The American Society for Cell Biology

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