Analysis on the physiological roles of phospholipases A_2

• Project/Area Number: 18580067
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied microbiology
• Research Institution: The University of Tokyo
• Principal Investigator: ARIOKA Manabu The University of Tokyo, Graduate School of Agricultural and Life Sciences, Associate Professor (20242159)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥3,800,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥300,000); Fiscal Year 2007: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2006: ¥2,500,000 (Direct Cost: ¥2,500,000)
• Keywords: phospholipase A_2 / lysophospholipid / MAP kinase / PC12 / Aspergillus oryzae / ホスホリパーゼA2 / nerve growth factor / MAP kinase / lysophosphatidylcholine / sphingosylphosphorylcholine / neurite outgrowth / secretory phospholipase A_2

Research Abstract

Nerve growth factor (NGF) induces MAP kinase activation and neuronal differentiation in PC12cells. We found that simultaneous addition of lysophospholipids, lysophosphatidylcholine (LPC) and sphingosylphosphorylcholine (SPC), markedly enhances NGF-induced MAP kinase activation, as examined by its phosphorylation, and neurite outgrowth. In agreement, the induction of immediate early gene expression by NGF was potentiated by LPC and SPC. Removal of extracellular Ca^<2+> did not show any effect. Studies using various inhibitors of suggested that the synergistic effect of NGF and LPC/SPC requires the activation of specific cellular signaling pathway(s).
Putative cytosolic phospholipase A_2 gene, AoplaA, uniquely found in Aspergillus oryzae genome was studied. Localization analysis using AoPlaA-EGFP fusion protein demonstrated that it localizes to mitochondria, which was supported by the biochemical fractionation experiment. To examine whether the N-terminal region contains the mitochondrial targeting signal, EGFP-fusion proteins were expressed and their localization was analyzed. The presence of cleavable mitochondrial targeting sequence was suggested by the N-terminal amino acid sequence analysis of AoPlaA-HA-His_6 purified on a Ni^<2+> column chromatography. In an independent experiment, a mutant strain ofA. olyzae disrupted for AoplaA was generated and tested for the phenotypic analyses.

Research Products

• [Journal Article] Lysophosphatidylcholine protects cerebellar granule neurons from apoptotic cell death. (2008)
  • Author(s): Ikeno, Y., Cheon, S. H., Konno, N., Nakamura, A., Kitamoto, K., Manabu Arioka
  • Journal Title: J. Neurosci. Res. (印刷中)(掲載確定)
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Lysophosphatidylcholine protects cerebellar granule neurons from apoptotic cell death. (2008)
  • Author(s): Ikeno, Y, Cheon, S.-h., Konno, N., Nakamura, A., Kitamoto, K., Arioka, M.
  • Journal Title: J. Neurosci. Res., (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Lysophosphatidylcholine protects cerebellar granule neurons from apoptotic cell death (2008)
  • Author(s): Ikeno, Y., Cheon, S. H., Konno, N., Nakamura, A., Kitamoto, K., Manabu Arioka
  • Journal Title: J. Neurosci. Res. (印刷中(掲載確定))
  • Peer Reviewed
• [Journal Article] Lysophosphatidylcholine mediates melanocyte dendricity through PKCzeta activation. (2007)
  • Author(s): Scott, G. A., Arioka, M., Jacobs, S. E.
  • Journal Title: J. Invest. Dermatol. 127 Pages: 668-675
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Novel neurotrophic effects of sphingosylphosphorylcholine in cerebellar granule neurons and in PC12 cells. (2007)
  • Author(s): Konno, N., Nakamura, A., Ikeno, Y., Cheon, S. H., Kitamoto, K., Arioka, M.
  • Journal Title: Biochem. Biophys. Res. Commun. 364 Pages: 163-168
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Lysophosphatidylcholine mediates melanocyte dendricity through PKCzeta activation. (2007)
  • Author(s): Scott, G. A., Arioka, M., Jacobs, S. E.
  • Journal Title: J. Invest. Dermatol. 127(3) Pages: 668-675
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Novel neurotrophic effects of sphingosylphosphorylcholine in cerebellar granule neurons and in PC12 cells. (2007)
  • Author(s): Konno, N., Nakamura, A., Ikeno, Y., Cheon, S.-h., Kitamoto, K., Arioka, M.
  • Journal Title: Biochem. Biophys. Res. Commun. 364(1) Pages: 163-168
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Novel neurotrophic effects of sphingosylphosphorylcholine in cerebellar granule neurons and in PC12 cells (2007)
  • Author(s): Konno, N., Nakamura, A., Ikeno, Y., Cheon, S. H., Kitamoto, K., Arioka, M.
  • Journal Title: Biochem. Biophys. Res. Commun. 364 Pages: 163-168
  • Peer Reviewed
• [Journal Article] Lysophosphatidylcholine mediates melanocyte dendricity through PKCzeta activation (2007)
  • Author(s): Scott, G. A., Arioka, M., Jacobs, S. E.
  • Journal Title: J. Invest. Dermatol. 127 Pages: 668-675
  • Peer Reviewed
• [Journal Article] Lysophosphatidylcholine mediates melanocyte dentricity through PKCζ activation (2007)
  • Author(s): Scott, G.A., Arioka, M., Jacobs, S.E.
  • Journal Title: J. Invest. Dermatol. 127・3 Pages: 668-675
• [Presentation] 神経成長因子NGFとリゾリン脂質の協調的作用の解析 (2008)
  • Author(s): 松木 友里, 北本 勝ひこ, 有岡 学
  • Organizer: 日本農芸化学会大会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-03-27
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Analysis on the synergistic neurotrophic effects of NGF and lysophospholipids (2008)
  • Author(s): Matsuki, Y, Kitamoto, K., Arioka, M.
  • Organizer: Annual Meeting of JSBBA
  • Place of Presentation: Nagoya, Japan
  • Year and Date: 2008-03-27
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 麹菌A. oryzaeの細胞質型ホスホリパーゼA_2の性質と機能 (2007)
  • Author(s): 高谷 康平, 北本 勝ひこ, 有岡 学
  • Organizer: 日本農芸化学会関東支部2007年度大会
  • Place of Presentation: 宇都宮
  • Year and Date: 2007-11-10
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] リゾリン脂質はNGFと協調的な神経栄養因子作用を示す (2007)
  • Author(s): 松木 友里, 有岡 学
  • Organizer: 日本農芸化学会関東支部2007年度大会
  • Place of Presentation: 宇都宮
  • Year and Date: 2007-11-10
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Characteristics of cytosolic phospholipase A_2 in A oryzae (2007)
  • Author(s): Takaya, K., Kitamoto, K., Arioka, M.
  • Organizer: Annual Meeting of JSBBA Kanto Branch
  • Place of Presentation: Utsunomiya, Japan
  • Year and Date: 2007-11-10
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] NGF and lysophospholipids demonstrate synergistic neurotrophic effects in PC12 cells (2007)
  • Author(s): Matsuki, Y., Arioka, M.
  • Organizer: Annual Meeting of JSBBA Kanto Branch
  • Place of Presentation: Utsunomiya, Japan
  • Year and Date: 2007-11-10
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 麹菌A. oryzae の細胞質型ホスホリパーゼA_2の性質と機能 (2007)
  • Author(s): 高谷康平, 北本勝ひこ, 有岡学
  • Organizer: 日本農芸化学会関東支部2007年度大会
  • Place of Presentation: 宇都宮
  • Year and Date: 2007-11-10
• [Presentation] Neurotrophic effects of secretory phospholipase A_2 and lysophospholipids (2007)
  • Author(s): Manabu Arioka
  • Organizer: 3rd International Conference on Phospholipases A_2 and Lipid Mediators
  • Place of Presentation: Sorrento, Italy
  • Year and Date: 2007-05-11
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 麹菌A. oryzaeにおける細胞質型ホスホリパーゼA_2の機能解析 (2007)
  • Author(s): 高谷 康平, 北本 勝ひこ, 有岡 学
  • Organizer: 日本農芸化学会大会
  • Place of Presentation: 東京
  • Year and Date: 2007-03-26
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Functional analysis of cytosolic phospholipase A_2 in A. oryzae (2007)
  • Author(s): Takaya, K., Kitamoto, K., Arioka, M.
  • Organizer: Annual Meeting of JSBBA
  • Place of Presentation: Tokyo, Japan
  • Year and Date: 2007-03-26
  • Description: 「研究成果報告書概要(欧文)」より