Analysis of lipid domains providing platforms for signal transduction

• Project/Area Number: 23590234
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General anatomy (including Histology/Embryology)
• Research Institution: Kagoshima University
• Principal Investigator: AKIKAZU Fujita 鹿児島大学, 共同獣医学部, 教授 (60282232)
• Project Period (FY): 2011-04-28 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2012: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2011: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 電子顕微鏡 / 脂質 / イノシトールリン脂質 / 脂質ラフト / 凍結活断レプリカ / 糖脂質 / ラフト / 急速凍結 / 凍結活断レプリカ法 / 超微形態 / 電子顕微鏡技術 / 情報伝達 / 凍結割断レプリカ / 超微細構造解析 / ホスホイノシトール / 生体膜 / レプリカ

Outline of Final Research Achievements

We have developed quick-freezing and freeze-fracture replica labeling (QF-FRL) method to analyze lipid distribution at the nano-meter level. The analysis of lipid distribution was limited to the lipids in the plasma membrane of the cell at first. In this study, we have further developed the QF-FRL method to analyze lipids in the membrane of various organelles at the inside of yeast and mammalian cells. And now we can establish the technique to specifically label phosphatidyl inositol 4-phosphat (PtdIns4P) by using anti-PtdIns4P antibody in the plasma membrane and the membrane of various organelle of inside cell. Now we examine the distribution of PtdIns4P at the nano-scale level.

Research Products

• [Journal Article] Clustering of Kir4.1 at specialized compartments of the lateral membrane in ependymal cells of rat brain (2015)
  • Author(s): Fujita A, Inanobe A, Hibino H, Nielsen S, Ottersen OP, Kurachi Y
  • Journal Title: Cell and Tissue Research Volume: 359(2) Issue: 2 Pages: 627-634
  • DOI: 10.1007/s00441-014-2030-6
  • Peer Reviewed
• [Journal Article] Yeast and mammalian autophagosomes exhibit distinct phosphatidylinositol 3-phosphate asymmetries (2014)
  • Author(s): Cheng J, Fujita A, Yamamoto H, Tatematsu T, Kakuta S, Obara K, Ohsumi Y, Fujimoto T.
  • Journal Title: Nat. Commun. Volume: 5 Issue: 1 Pages: 3207-3207
  • DOI: 10.1038/ncomms4207
  • Peer Reviewed / Open Access
• [Journal Article] A method for efficient observation of intracellular membranes of monolayer culture cells by quick-freeze and freeze-fracture electron microscopy. (2012)
  • Author(s): Akikazu Fujita, Toyoshi Fujimoto, Nami Ozato-Sakurai, Hitomi Suzuki
  • Journal Title: J Electron Microsc (Tokyo) Volume: 61 Issue: 6 Pages: 441-446
  • DOI: 10.1093/jmicro/dfs063
  • Peer Reviewed
• [Journal Article] The distribution of phosphatidylinositol 4,5-bisphosphate in acinar cells of rat pancreas revealed with the freeze-fracture replica labeling method (2011)
  • Author(s): Nami Ozato-Sakurai, Akikazu Fujita, Toyoshi Fujimoto
  • Journal Title: PLoS One Volume: 6
  • Peer Reviewed
• [Presentation] 免疫電顕法を用いたホスホイノシタイドの微細局在 (2013)
  • Author(s): 藤田秋一、程晶磊、藤本豊士
  • Organizer: 日本平滑筋学会
  • Place of Presentation: 旭川
  • Invited
• [Presentation] ラット膵臓外分泌細胞におけるPI(4,5)P2の微細分布 (2011)
  • Author(s): 藤田秋一、尾里ー櫻井納美、藤本豊士
  • Organizer: 日本解剖学会 第67回九州支部学術集会
  • Place of Presentation: 宮崎県宮崎市
• [Book] 獣医生化学 (2015)
  • Author(s): 藤田 秋一
  • Publisher: 朝倉書店
• [Book] 獣医生化学 (2015)
  • Author(s): 藤田秋一
  • Total Pages: 10
  • Publisher: 朝倉書店
• [Remarks] 電子顕微鏡による脂質分布のナノスケール解析
  • URL: http://www.vet.kagoshima-u.ac.jp/kadai/V-Mol/bunshibyoutai01/Research.html