Fish hematogenesis function associated with defense system to pathogens

• Project/Area Number: 24658176
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: General fisheries
• Research Institution: Nippon Veterinary and Life Science University
• Principal Investigator: KURATA Osamu 日本獣医生命科学大学, 獣医学部, 准教授 (90277666)
• Co-Investigator(Kenkyū-buntansha): NAKAYASU Chihaya 独立行政法人水産総合研究センター, その他 (00311225) ; OKAUCHI Masanori 独立行政法人水産総合研究センター, 研究員 (40372023) ; OZAKI Akiyuki 独立行政法人水産総合研究センター, 研究員 (40416045)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2014: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 造血細胞 / 移植 / 細胞分化 / 細胞表面マーカー / 遺伝子導入 / 魚類 / ヒラメ / 細胞・組織 / 免疫学 / 発生・分化 / 造血

Outline of Final Research Achievements

We aimed to analyze fish hematogenesis function by utilizing a hematopoietic precursor cell line established from Japanese flounder. We were successful in transplantation of the hematopoietic precursor cells (HPC) into fish with the same genetic lineage and larvae of flounder, Transplanted cells were detected in a hematopoietic organ (kidney) of recipient fish. Some monoclonal antibodies to cell surface molecules were established for estimation of differentiation in Japanese flounder hematopoietic cells. It was possible to recognize immature or mature cells using combinations of the antibodies. We tried to establish GFP-expressing HPC in order to track transplanted cells. Unfortunately, we could not achieve it but suitable expression vector and gene transfection method was found. As a new knowledge, cell division in the hematopoietic organ was induced 2 to 3 days after administration of pathogens.

Research Products

• [Journal Article] Ontogeny of anti-viral hemorrhagic septicemia virus (VHSV) immunity in developing Japaneseflounder. (2012)
  • Author(s): Matsuyama, T., Nakayasu, C., Fujiwara,A., Kurita, Takano, T., Ito, T. and Sano, M.
  • Journal Title: Dev. Comp. Immunol. Volume: 37 Issue: 3-4 Pages: 312-322
  • DOI: 10.1016/j.dci.2012.02.014
  • Peer Reviewed
• [Presentation] N末領域を欠くヒラメIL-8類似分子について (2015)
  • Author(s): 倉田 修、萩原文乃、和田新平、松山知正、坂井貴光
  • Organizer: 平成27年度日本水産学会春季大会
  • Place of Presentation: 東京海洋大学
  • Year and Date: 2015-03-28
• [Presentation] Edwardsiella tarda感染ヒラメの肝臓におけるインターロイキン8の局在 (2015)
  • Author(s): 倉田 修、萩原文乃、和田新平
  • Organizer: 平成27年度日本魚病学会春季大会
  • Place of Presentation: 東京海洋大学
  • Year and Date: 2015-03-08
• [Presentation] Inhibition of chemotactic activity of flounder IL-8 by specific antibodies for its N-terminal region
  • Author(s): Kurata O, Matsuyama T, Sakai T, Takano T, Wada S, Nakasyasu C
  • Organizer: 1st International Conference of Fish and Shellfish Immunology
  • Place of Presentation: Vigo, Spain
• [Presentation] ヒラメEF‐1αプロモーター制御によるGFP発現コンストラクトの作製
  • Author(s): 倉田 修、和田新平
  • Organizer: 平成26年度日本水産学会春季大会
  • Place of Presentation: 北海道大学函館キャンパス
• [Presentation] Edwardsiella tarda感染ヒラメにおける肝臓の初期病理変化
  • Author(s): 萩原文乃、倉田 修、和田新平
  • Organizer: 平成26年度日本魚病学会春季大会
  • Place of Presentation: 函館国際ホテル
• [Presentation] 好中球の走化性に関与するヒラメIL-8のN末領域
  • Author(s): 倉田 修、和田新平、松山知正、坂井貴光、高野倫一
  • Organizer: 平成24年度日本水産学会秋季大会
  • Place of Presentation: 独立行政法人水産大学校
• [Presentation] ヒラメIL-8タンパク検出のための特異抗体の作製
  • Author(s): 倉田 修、和田新平、松山知正、坂井貴光、高野倫一、中易千早
  • Organizer: 平成25年度日本水産学会春季大会
  • Place of Presentation: 東京海洋大学（品川キャンパス）