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Completion of Meiosis Is Not Always Required for Acrosome Formation in HSP70-2 Null Mice¹

^a Department of Anatomy and Developmental Biology and Central Laboratory for Electron Microscopy, ^b Faculty of Medicine, Kyoto University, Kyoto 606-8501, Japan ^c Gamete Biology Section, Laboratory of Reproductive and Developmental Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina 27709 ^d Environmental Carcinogenesis Division, Reproductive Toxicology Division, ^e National Health and Environmental Effects Research Laboratory, US Environmental Protection Agency, Research Triangle Park, North Carolina 27711 ^f Department of Anatomy, Miyazaki Medical College, Miyazaki 889-16, Japan

Hsp70-2 is a unique member of the mouse 70-kDa heat shock protein family that is synthesized during meiosis in spermatogenic cells. Germ cells in male mice homozygous for a targeted mutation in the Hsp70-2 gene (Hsp70-2^-/-) arrest in development and undergo apoptosis at the end of the pachytene spermatocyte stage of meiotic prophase. However, cells with a putative acrosome were present occasionally in histological sections of the testes of juvenile and adult Hsp70-2^-/- mice. This study verified that acrosomes were present and investigated the relationship between acrosome formation and the process of meiosis. Histochemistry with the periodic acid-Schiff procedure and immunostaining with monoclonal antibody MN7 verified that acrosomes were present in Hsp70-2^-/- mice, and electron microscopy showed that some of these cells had condensing nuclei characteristic of step 8–9 spermatids. The frequency of acrosome-containing cells in Hsp70-2^-/- mice was less than 0.01% of that in wild-type mice. Propidium iodide staining and cytophotometry indicated that the average DNA content of nuclei in MN7-positive cells in Hsp70-2^-/- mice was usually about twice, or occasionally the same as, that of nuclei in round spermatids of wild-type mice. Meiotic metaphase I and II chromosome spreads were observed in spermatogenic cells from Hsp70-2^-/- mice but at a much lower frequency than in wild-type mice. These results indicate that not all pachytene spermatocytes in Hsp70-2^-/- mice arrest in meiosis, but they may divide once or sometimes twice and begin acrosome formation and nuclear condensation. This demonstrates that some aspects of spermatid development can occur without the completion of meiosis in mice, as has been reported recently for Drosophila.

¹ This work was supported by grants from the Japanese Ministry of Education, Science and Culture, and the Toyota Foundation.

² Correspondence. FAX: 81 75 751 7529; mori{at}med.kyoto-u.ac.jp

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