Elucidation of the dynamics and functions of lipid rafts in the filopodia formation by simultaneous visualization of lipid distribution and membrane-lipid phase

• Project/Area Number: 24770135
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Functional biochemistry
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: KISHIMOTO Takuma 独立行政法人理化学研究所, 小林脂質生物学研究室, 基礎科学特別研究員 (70585158)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2012: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: 脂質ドメイン / 細胞膜二重膜 / 膜物性 / 脂質分布 / アクチン細胞骨格 / 糸状仮足 / 低分子量Gタンパク質 / 脂質輸送分子 / コレステロール / Cdc42 / 細胞膜 / 脂質二重層 / 細胞二重膜 / 細胞形態

Research Abstract

Elucidating the machineries and dynamics of lipid rafts and the correlations with cytoskeleton on the plasma membrane is of crucial importance for determining mechanisms underlying cell events. In this study, we employed total internal reflection fluorescence microscopy to precisely visualize lipids of the plasma membrane and examined dynamics of lipids during HeLa cells spreading. We found that the distribution of these raft lipids segregated progressively and that lipids form separated domains in the outer leaflet of the plasma membrane during cell spreading. Cholesterol domains polarized at the tip of filopodia-like protrusion with liquid ordered phase. In addition, our results suggest the possibility that these domains are formed dynamically via feedback regulation mechanisms involving signal lipid PIP2 domains, small GTPase Cdc42, and ATP binding cassette transporter ABCA1 regulated by Cdc42.

Research Products

• [Journal Article] Binding of a pleurotolysin ortholog from Pleurotus eryngii to sphingomyelin and cholesterol-rich membrane domains. (2013)
  • Author(s): Bhat H B, Kishimoto T, Abe M, Makino A, Inaba T, Murate M, Dohmae N, Kurahashi A, Nishibori K, Fujimori F, Greimel P, Ishitsuka R, Kobayashi T.
  • Journal Title: Journal of Lipid Research Volume: 54 Issue: 10 Pages: 2933-2943
  • DOI: 10.1194/jlr.d041731
  • Peer Reviewed
• [Journal Article] 脳の脂質と疾患 (2012)
  • Author(s): 岸本拓磨、稲葉岳彦、酒井翔太、小林敏秀
  • Journal Title: 最新精神医学 Volume: 51 17巻3号 Pages: 765-775
• [Presentation] 細胞膜ラフトにおけるコレステロールドメインの脂質分布とアクチン細胞骨格の相関 (2013)
  • Author(s): 岸本拓磨、小林俊秀
  • Organizer: 第36回分子生物学会
  • Place of Presentation: 神戸
  • Year and Date: 2013-12-03
• [Presentation] Membrane deformation by phospholipase C beta 1 (2013)
  • Author(s): Kishimoto, Takuma, Inaba, Takehiko, Tajima, Takuya, Abe, Mitsuhiro, Murate, Motohide, Makino, Asami, Ishitsuka, Reiko, Ikeda, Yasuo, Takeoka, Shinji, Kobayashi, Toshihide
  • Organizer: Lipid-protein interaction : From molecules to cells
  • Place of Presentation: Wako, Japan
  • Year and Date: 2013-05-09
• [Presentation] Binding of pleurotolysin A ortholog from Pleurotus eryngii to sphingomyelin and cholesterol-rich membrane domains (2013)
  • Author(s): Balakrishna BH, Kishimoto Takuma, Abe Mitsuhiro, Makino Asami, Dohmae Naoshi, Kurahashi Atsushi, Nishibori Kozo, Fujimori Fumihiro, Ishitsuka Reiko, and Kobayashi Toshihide
  • Organizer: International Forum for Membrane Research
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2013-03-16
• [Presentation] Transformation of phosphatidyl- ethanolamine- and phosphatidylserine -containing membranes by phospholipase C beta 1 (2013)
  • Author(s): Tajima Takuya, Inaba Takehiko, Kishimoto Takuma, Abe Mitsuhiro, Murate Motohide, Makino Asami, Sakai Shota, Hullin-Matsuda Francoise, Ishitsuka Reiko,Ikeda Yasuo, Takeoka Shinji, and Kobayashi Toshihide
  • Organizer: International Forum for Membrane Research
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2013-03-16
• [Presentation] 細胞膜ラフトにおけるコレステロールドメインの脂質分布とアクチン細胞骨格の相関 (2013)
  • Author(s): 岸本拓磨、小林俊秀
  • Organizer: 分子生物学会
  • Place of Presentation: 神戸
• [Presentation] Phospholipase C-beta 1b changes the curvature of phosphatidylethanolamine-containing membranes (2012)
  • Author(s): Tajima Takuya, Inaba Takehiko, Kishimoto Takuma, Abe Mitsuhiro, Murate Motohide, Sakai Shota, Ishitsuka Reiko, Takeoka Shinji, and Kobayashi Toshihide
  • Organizer: Lipid-Protein interactions in Membranes : Implications for Health and Disease
  • Place of Presentation: Hyderabad, India
  • Year and Date: 2012-11-02