Improvement of ICSI technique focused on the induction of oocyte activation in pigs

• Project/Area Number: 26850172
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Animal production science
• Research Institution: National Agriculture and Food Research Organization (2016); National Institute of Agrobiological Sciences (2014-2015)
• Principal Investigator: NAKAI Michiko 国立研究開発法人農業・食品産業技術総合研究機構, 生物機能利用研究部門 動物機能利用研究領域, 研究員 (30442825)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: 顕微授精 / ブタ / 卵活性化 / PLCzeta / ICSI / 受精

Outline of Final Research Achievements

Intracytoplasmic sperm injection (ICSI) is a useful tool for producing live offspring from immotile sperm. However, the efficiency of fertilization after ICSI is low. It is hypothesized that a failure in oocyte activation causes it in this study. Therefore, this study tried to develop new method for induction of physiological oocyte activation using PLCzeta, which is thought to be the oocyte-activating factor in mammalian sperm. As a result, it was clearly that a lack of PLCzeta in sperm led to a failure in an induction of oocyte activation after ICSI. Furthermore, the injection of PLCzeta mRNA into the pig oocyte injected with sperm induced oocyte activation efficiently and promoted dramatically fertilization. In addition, the induction of oocyte activation using PLCzeta protein will be continued to investigate.

Research Products

• [Journal Article] Efficient pig ICSI using Percoll-selected spermatozoa; evidence for the essential role of phospholipase C-ζ in ICSI success (2016)
  • Author(s): Michiko NAKAI, Shun-ichi SUZUKI, Junya ITO, Dai-ichiro FUCHIMOTO, Shoichiro SEMBON, Junko NOGUCHI, Akira ONISHI, Naomi KASHIWAZAKI, Kazuhiro KIKUCHI
  • Journal Title: Journal of Reproduction and Development Volume: 62 Issue: 6 Pages: 639-643
  • DOI: 10.1262/jrd.2016-103
  • NAID: 130005251190
  • ISSN: 0916-8818, 1348-4400
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Lack of calcium oscillation causes failure of oocyte activation after intracytoplasmic sperm injection in pigs. (2016)
  • Author(s): Nakai M, Ito J, Suzuki S, Fuchimoto D, Sembon S, Suzuki M, Noguchi J, Kaneko H, Onishi A, Kashiwazaki N, Kikuchi K
  • Journal Title: Journal of Reproduction and Development Volume: 62 Issue: 6 Pages: 615-621
  • DOI: 10.1262/jrd.2016-113
  • NAID: 130005251188
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] 卵細胞質内精子注入(ICSI)後のブタ卵における卵活性化不全の原因究明 (2016)
  • Author(s): 中井美智子， 伊藤潤哉， 鈴木俊一， 淵本大一郎， 千本正一郎， 野口純子， 金子浩之，柏崎直巳， 大西彰，菊地和弘
  • Organizer: 日本繁殖生物学会
  • Place of Presentation: 麻布大学 (神奈川県 相模原市)
  • Year and Date: 2016-09-11
• [Presentation] Lack of phospholipase Czeta causes fertilization failure after intracytoplasmic sperm injection in pigs (2016)
  • Author(s): Nakai M, Suzuki S, Ito J, Fuchimoto D, Sembon S, Noguchi J, Kashiwazaki N, Kikuchi K
  • Organizer: 第18回 International Congress on Animal Reproduction
  • Place of Presentation: フランス (Tours)
  • Year and Date: 2016-06-27
  • Int'l Joint Research
• [Presentation] Lack of phospholipase C causes fertilization failure after intracytoplasmic sperm injection in pigs (2016)
  • Author(s): Nakai M, Suzuki S, Ito J, Fuchimoto D, Sembon S, Noguchi J, Kashiwazaki N, Kikuchi K
  • Organizer: 第18回International Congress on Animal Reproduction
  • Place of Presentation: フランス, トゥール
  • Year and Date: 2016-06-26
  • Int'l Joint Research