Application of convenience gender-sorting of mouse spermatozoa to mouse reproductive engineering technology

2011 Fiscal Year Final Research Report

• Project/Area Number: 22650095
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Laboratory animal science
• Research Institution: 実験動物中央研究所 (2011); Central Institute for Experimental Animals (2010)
• 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)
• Project Period (FY): 2010 – 2011
• Keywords: 動物実験技術 / 体外受精 / 胚移植 / 雌雄生み分け

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.

Research Products

• [Journal Article] Study on establishment of congenic strains and screening of characteristics in IRS-2 deficient mice to support translational research on type 2 diabetes (2011)
  • Author(s): Haruo HASHIMOTO
  • Journal Title: Experimental animals Volume: 60巻 Pages: 21-32
  • DOI: DOI:10.1538/expanim.60.21
  • Peer Reviewed
• [Journal Article] db/dbマウスの系統比較から考察されるレジスチンの役割 (2011)
  • Author(s): 橋本晴夫、新井敏郎、川井健司、鬼頭千佳、大美典子、早川絵里、水島友子、樋口裕一郎、江袋美知、末水洋志、日置恭司、斎藤宗雄、伊藤守、小坂樹徳
  • Journal Title: 実験動物技術 Volume: 46巻 Pages: 11-18
  • URL: http://ci.nii.ac.jp/naid/10029381001
  • Peer Reviewed
• [Journal Article] Vinyl isolator breeding induces insulin resistance in C57BL/6JJcl mice (2011)
  • Author(s): Haruo HASHIMOTO, Susumu EBUKURO, Ryoko NOZU, Masami UENO, Toshiro ARAI, Kenji KAWAI, Hiroshi HIRATA, Maya OGAWA, Takuma MIZUSAWA, Kuniyasu IMAI, Yuichiro HIGUCHI, Hiroshi SUEMIZU, Mamoru ITO, Muneo SAITO, and Kyoji HIOKI
  • Journal Title: Experimental animals Volume: 60巻 Pages: 497-508
  • DOI: DOI:10.1538/expanim.60.497
  • Peer Reviewed
• [Presentation] アイソレーター飼育はC57BL/6JJclマウスにインシュリン抵抗性を引き起こす (2011)
  • Author(s): 橋本晴夫、江袋進、野津量子、植野昌未、新井敏郎、川井健司、平田裕、小川麻耶、水澤卓馬、末水洋志、伊藤守、齊藤宗雄、日置恭司(実中研,日獣大)
  • Organizer: 第58回日本実験動物学会総会
  • Place of Presentation: 東京
  • Year and Date: 2011-05-26