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Mol. Hum. Reprod. Advance Access published online on February 11, 2005
Molecular Human Reproduction, doi:10.1093/molehr/gah136
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Molecular Human Reproduction © European Society of Human Reproduction and Embryology 2004; all rights reserved
Received January 7, 2004
Revised November 11, 2004
Accepted November 15, 2004

Article

Expression of Smac/DIABLO in mouse preimplantation embryos and its correlation to apoptosis and fragmentation

Yoko Honda 1, Hideo Tanikawa 1, Jun Fukuda 1*, Kazuhiro Kawamura 1, Naoki Sato 1, Toshiharu Sato 1, Yasushi Shimizu 1, Hideya Kodama 1, and Toshinobu Tanaka 1

1 Division of Obstetrics and Gynecology, Department of Reproductive and Developmental Medicine, Akita University School of Medicine, 1-1-1, Hondo, Akita 010-8543, Japan

* To whom correspondence should be addressed.
Jun Fukuda, E-mail: fukudaj{at}obgyn.med.akita-u.ac.jp


  Abstract

Regulation of early embryonal development during fertilization and implantation is crucial for mammalian reproduction. Several studies have described cell death during preimplantation embryogenesis in a range of mammalian species, both in vivo and in vitro. Therefore, apoptosis may be involved in early embryonic arrest and the characteristic cytoplasmic fragments are the equivalents of apoptotic bodies, the end-product of apoptosis. Although apoptosis is expected to associate with fragmentation in early preimplantation embryos, the mechanism through which this fragmentation occurs has not been elucidated. Recently, second mitochondria-derived activator of caspase/Direct IAP Binding Protein with Low pI (Smac/DIABLO) was identified as a mitochondrial protein that is released into the cytosol during apoptosis. Once released, the Smac/DIABLO blocks the anti-apoptotic activity of inhibitor of apoptosis proteins (IAPs). We hypothesized that the Smac/DIABLO may be involved in the fragmentation of mouse preimplantation embryos. Therefore, we investigated the expression of Smac/DIABLO mRNA and protein and its localization in mouse oocytes and preimplantation embryos. Smac/DIABLO mRNA was detected by RT-PCR in the oocytes and the preimplantation embryos. Immunohistochemistry studies showed that the Smac/DIABLO protein localized in mitochondria and was released into the cytosol in both fragmented embryos and embryos in which apoptosis was induced by staurosporine. These observations indicate that the Smac/DIABLO is involved in the fragmentation and apoptosis of preimplantation embryos.

Keywords: apoptosis; preimplantation embryo; Smac/DIABLO.
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