| Project/Area Number |
22650095
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Laboratory animal science
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| Research Institution | 実験動物中央研究所 (2011)
Central Institute for Experimental Animals (2010) |
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Principal Investigator |
HASHIMOTO Haruo 実験動物中央研究所, バイオメディカル研究部, 研究員 (30353478)
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| Project Period (FY) |
2010 – 2011
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| Project Status |
Completed (Fiscal Year 2011)
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| Budget Amount *help |
¥3,290,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥390,000)
Fiscal Year 2011: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2010: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | 動物実験技術 / 体外受精 / 胚移植 / 雌雄生み分け / マウス / 雌雄産み分け / SRY / 雌雄生みわけ / Sry遺伝子 |
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| Research Abstract |
In this study, we attempted to apply convenience gender sorting methods for the mammals to mice with Cy3 labeling for intracytoplasmic sperm injection(ICSI) and magnetic beads for in vitro fertilization(IVF), via sex-determining region Y(SRY) antibody.
The Cy3-SRY antibody conjugate was added to TYH medium suspended mouse spermatozoa. The mouse spermatozoa labeled with Cy3-SRY antibody conjugate were used to ICSI. On the other hand, after reaction of the SRY antibody and mouse spermatozoa in TYH medium, magnetic beads were added and incubated. Thereafter, the conjugates of spermatozoa, SRY antibody and magnetic beads were pulled to the bottom of the medium by a magnet and the supernatant in the medium was used for IVF. The embryos were transferred to pseudo-pregnant mice, and the gender ratio was investigated on day 18 of gestation by Cesarean section.
As these results, the ICSI using the spermatozoa with Cy3-SRY antibody conjugate succeeded to reproduce 31 males of 36 infants(86.1%). On the other hand, the female proportion reproduced by the IVF using the spermatozoa with conjugates of SRY antibody and magnetic beads was 67.3%.
It was considered that these methods are very effective in reproduction of transgenic mice with more than 2 mutation genes that require several times of IVF and ET.
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