Demethylation Dynamics of the Paternal Genome in Pronuclear-Stage Bovine Zygotes Produced by In Vitro Fertilization and Ooplasmic Injection of Freeze-Thawed or Freeze-Dried Spermatozoa
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概要
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This study was designed to investigate the dynamics of the paternal genome demethylation in pronuclear-stage bovine zygotes produced either by in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) using freeze-thawed (FT) as well as freeze-dried (FD) bull sperm stored at +4 or -196 C for one year. Zygotes were fixed and immunostained using anti-5-methyl-cytosin at 8, 10, 14 and 18 h post IVF (hpi) and at 6 and 12 h post ICSI (hpic). In conventional IVF-derived zygotes, the overall average of the relative methylation (RM; male/female) decreased from 0.92 at 8 hpi to 0.69 at 10 hpi (P<0.05) without any additional decrease at 14 and 18 hpi (0.67 and 0.64, respectively; P>0.05). This was accompanied by higher proportions of zygotes showing RM<0.6 (45.5, 37.5 and 38.2% at 10, 14 and 18 hpi, respectively; P<0.05) compared with 3.7% at 8 hpi. The overall averages of the RM in the FT-ICSI derived zygotes (0.79 and 0.66 at 6 and 12 hpic, respectively) were similar to those in the corresponding IVF-derived zygotes (8 and 14 hpi), but a higher proportion of the 6 hpic zygotes (37.8%; P<0.05) showed an RM<0.6 compared with the 8 hpi zygotes (3.7%). The proportions of FD-ICSI derived zygotes at 12 hpic showing an RM<0.6 (60.6 and 62.4% for +4 and -196 C storage, respectively) were higher than that of the FT-ICSI derived zygotes (39.4%; P<0.05). Thus, the bovine paternal genome rapidly demethylated within 10 h after IVF and 6 h after ICSI, and the freeze-drying and/or the storage process had no adverse effect on demethylation of the paternal genome. The extent of demethylation in the pronuclear-stage bovine zygotes was moderate, with 0.4≤RM<0.6.
- 日本繁殖生物学会の論文
- 2009-08-01
著者
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HIRABAYASHI Masumi
National Institute for Physiological Sciences
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Hirabayashi Masumi
信州大学 繊維
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ABDALLA Hany
Interdisciplinary Graduate School of Science and Technology, Shinshu University
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HOCHI Shinichi
Interdisciplinary Graduate School of Science and Technology, Shinshu University
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ABDALLA Hany
Graduate School of Science and Technology, Shinshu University
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HOCHI Shinichi
Graduate School of Science and Technology, Shinshu University
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Abdalla Hany
Interdisciplinary Graduate School Of Science And Technology Shinshu University
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Hochi Shinichi
Fac. Of Textile Sci. And Technol. Shinshu Univ.
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Hochi Shinichi
Interdisciplinary Graduate School Of Science And Technology Shinshu University
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Hirao Masao
Ina Research Laboratory Kitayama Labes Co. Ltd.
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Hochi Shinichi
Graduate School Of Science And Technology Shinshu University
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Abdalla Hany
Graduate School Of Science And Technology Shinshu University
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Yoshizawa Yusuke
National Institute for Physiological Sciences
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