細胞外ATPを介したアストログリア-ニューロン相互調節機構の解明

• Project/Area Number: 15082212
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Kyushu University (2004-2007); National Institute of Health Sciences (2003)
• Principal Investigator: 井上 和秀 Kyushu University, 大学院・薬学研究院, 教授 (80124379)
• Co-Investigator(Kenkyū-buntansha): 津田 誠 九州大学, 大学院・薬学研究院, 准教授 (40373394) ; 小泉 修一 山梨大学, 大学院・医学工学総合研究部, 教授 (10280752)
• Project Period (FY): 2003 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥59,500,000 (Direct Cost: ¥59,500,000); Fiscal Year 2007: ¥11,900,000 (Direct Cost: ¥11,900,000); Fiscal Year 2006: ¥11,900,000 (Direct Cost: ¥11,900,000); Fiscal Year 2005: ¥11,900,000 (Direct Cost: ¥11,900,000); Fiscal Year 2004: ¥11,900,000 (Direct Cost: ¥11,900,000); Fiscal Year 2003: ¥11,900,000 (Direct Cost: ¥11,900,000)
• Keywords: ATP / アストロサイト / 細胞死 / 活性酸素 / MAPK / 保護作用 / ATP受容体 / P2Y1 / 神経保護作用 / H2O2 / DNAチップ解析 / 酸化ストレス / 神経細胞死 / グリオトランスミッション / カルシウムウエーブ / 三者間シナプス / シナプス伝達 / 海馬

Research Abstract

本研究では、これまでに次のことを明らかにした。(1)アストロサイトにおいて酸化ストレス緩和に関与する遺伝子群(oxidoreductase genes)の発現をATPが上昇させた。(2)ATPは、酸化ストレス過酸化水素誘発アストロサイト細胞死をP2Yl受容体を介して抑制した。(3)過酸化水素誘発アストロサイト細胞死の詳細な機序を明らかにした。(4)金属イオン依存性蛋白質分解酵素〜9(MMP-9)は細胞外マトリックスを除去し、細胞の増殖、突起進展の際に空間を提供する非常に重要な分子であるが、一方で種々の細胞外分子を分解するために、BBB破壊等の細胞外環境破壊も引き起こす。ATPはアストロサイトのTNF-alpha産生能をtonicに抑制することにより、MMP-9産生量を調節し、脳内恒常性の維持に関与しているものと考えられた。本年度は、このようなアストロサイト自身に対する保護効果が、過酸化水素の非常に強い神経細胞障害性に対してどのような影響を及ぼすのかを検討した。その結果、gliotransmitterであるATPがP2Yl受容体活性化を介したアストログリアの細胞死回避・機能維持により、実際に酸化ストレス誘発性の神経細胞死を防御することを明らかにした。さらに、アストログリア細胞のP2Y1受容体活性化によりサイトカインIL-6の放出が促進され、ATP刺激アストログリアの神経保護効果がIL-6中和抗体共添加により抑制されたこと,さらに外因的に加えたIL-6がH202誘発障害に対して神経保護作用を示すことから、ATPはアストログリアからのIL-6産生・放出機能を促進することにより神経細胞を保護することを明らかにした。このように、これまでの脳保護薬とは異なったアストログリアのATP受容体をターゲットとした全く新規の創薬の一助となる可能性がある。

Research Products

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• [Journal Article] Fibronectin/integrin system is involved in P2X4 receptor upregulation in the spinal cord and nueropathic pain aftervnerve injury. (2008)
  • Author(s): M., Tsuda,E., Toyomitsu,T., Komatsu,T., Masuda,E., Kunifusa,K., Nasu-Tada,S., Koizumi,K., Yamamoto,J., Ando,K., Inoue
  • Journal Title: Glia (In press)
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• [Journal Article] Y. Shinozaki, Y. Sato, S. Koizumi, Y. Ohno, T. Nagao and K. Inoue. (2007)
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• [Journal Article] Reduced pain behaviors and ERK activation in primary sensory neuros by peripheral tissue injury in mice lacking platelet-activating factor receptor. (2007)
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• [Journal Article] Involvment of P2X(4) and P2Y(12) receptors in ATP-induced microglial chemotaxis. (2007)
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• [Journal Article] UDP facilitates microglial phagocytosis through P2Y6 receptors. (2007)
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  • Journal Title: Cell Adhesion and Migration 1 Pages: 131-132
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• [Journal Article] Modification of neuropathic pain sensation through microglial ATP receptors (2007)
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• [Journal Article] 神経因性疼痛におけるミクログリアの関与 (2007)
  • Author(s): 井上 和秀
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• [Journal Article] ATP受容体を介する慢性疼痛発症機構 (2007)
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• [Journal Article] 神経因性疼痛と脊髄ミクログリアのATP受容体 (2007)
  • Author(s): 津田誠, 井上和秀
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  • NAID: 40015625659
• [Journal Article] 基礎と臨床のリレーショナル疼痛研究:神経因性疼痛におけるミクログリアと P2X4 受容体の関与 (2007)
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  • Journal Title: Pain Research 22 Pages: 163-169
  • NAID: 130004651885
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• [Journal Article] 神経因性疼痛におけるミクログリアとATP受容体の関与 (2007)
  • Author(s): 井上 和秀
  • Journal Title: 医学の歩み 223 Pages: 681-686
• [Journal Article] NMDA receptor-independent synaptic plasticity in the central amygdala in the rat model of neuropathic pain. (2007)
  • Author(s): Ikeda R, Takahashi Y, Inoue K, Kato F
  • Journal Title: Pain 127 Pages: 161-172
• [Journal Article] Involvement of P2X(4) and P2Y(12) receptors in ATP-induced microglial chemotaxis. (2007)
  • Author(s): Ohsawa K, Irino Y, Nakamura Y, Akazawa C, Inoue K, Kohsaka S
  • Journal Title: Glia 55 Pages: 604-616
• [Journal Article] Long-term potentiation of neuronal excitation by neuron-glia interactions in the rat spinal dorsal horn. (2007)
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  • Journal Title: Eur J Neurosci (In press)
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• [Journal Article] UDP acting at P2Y6 receptors is a mediator of microglial phagocytosis. (2007)
  • Author(s): S.Koizumi, Y.Shigemoto-Mogami, K.Nasu-Tada, Y.Shinozaki, K.Ohsawa, M.Tsuda, B.V.Joshi, K.A.Jacobson, S.Kohsaka, K.Inoue
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• [Journal Article] Possible involvement of increase in spinal fibronectin following peripheral nerve injury in upregulation of microglial P2X4, a key molecule for mechanical allodynia. (2006)
  • Author(s): Nasu-Tada K., Koizumi S., Tsuda M., Kunifusa E., Inoue K
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• [Journal Article] Direct excitation of deep dorsal horn neurones in the rat spinal cord by the activation of postsynaptic P2X receptors. (2006)
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• [Journal Article] Retinoic acids increase P2X2 receptor expression through the 5'-flanking region of P2rx2 gene in rat phaeochromocytoma PC-12 cells. (2006)
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• [Journal Article] Extracellular ATP counteracts the ERK1/2-mediated death-promoting signaling cascades in astrocytes. (2006)
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• [Journal Article] Microglial・7 nicotinic acetylcholine receptors drive a phospholipase C/IP3 pathway and modulate the cell activation toward a neuroprotective role. (2006)
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• [Journal Article] Screening of novel nuclear receptor agonists by a convenient reporter gene assay system using green fluorescent protein derivatives. (2006)
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• [Journal Article] Involvement of β1 integrin in microglial chemotaxis and proliferation on fibronectin : different regulations by ADP through PKA. (2005)
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• [Journal Article] Cytoprotection against oxidative stress-induced damage of astrocytes by extracellular ATP via P2Y1 receptors. (2005)
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• [Journal Article] Neuropathic pain and spinal microglia : a big problem from molecules in ‘small'glia. (2005)
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• [Journal Article] Activation of p38MAPK in spinal hyperactive microglia contributes to neuropathic pain hypersensitivity following peripheral nerve injury. (2004)
  • Author(s): M.Tsuda, A.Mizokoshi, Y.Shigemoto-Mogami, S.Koizumi, K.Inoue
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• [Journal Article] Production and release of neuroprotective TNF by P2X7 receptor-activated microglia. (2004)
  • Author(s): T.Suzuki, I.Hide, K.Ido, A.Inoue, S.Kohsaka, K.Inoue, Y.Nakata
  • Journal Title: J.Neurosci. 24 Pages: 1-7
• [Journal Article] Ca^<2+> waves in keratinocytes are transmitted to sensory neurons : the involvement of extracellular ATP and P2Y2 receptor activation. (2004)
  • Author(s): S.Koizumi, K.Fujishita, K.Inoue, Y.Shigemoto-Mogami, M.Tsuda, K.Inoue
  • Journal Title: Biochem.J. 380 Pages: 329-338
• [Journal Article] Differential modulation of PI3-kinase/Akt pathway during all-trans retinoic acid- and Am80-induced HL-60 cell differentiation revealed by DNA microarray analysis. (2004)
  • Author(s): S.Ishida, Y.Shigemoto-Mogami, Y.Shinozaki, H.Kagechika, K.Shudo, S.OzaWa, J.Sawada, Y.Ohno1, K.Inoue
  • Journal Title: Biochemical Pharmacology 68 Pages: 2177-2186
• [Journal Article] Cytoprotection against oxidative-stress-induced damage of astrocytes by extracellular ATP via P2Y1 receptors. (2004)
  • Author(s): Y.Shinozaki, S.Koizumi, S.Ishida, J.Sawada, Y.Ohno, K.Inoue
  • Journal Title: Glia 49 Pages: 288-300
• [Journal Article] Direct excitation of inhibitory interneurons by extracellular ATP mediated by P2Y1 receptors in the hippocampal slice. (2004)
  • Author(s): M.Kawamura, C.Gachet, K.Inoue, F.Kato
  • Journal Title: J.Neurosci. 24 Pages: 10835-10845
• [Presentation] UDP, P2Y6 and microglial phagocytosis.(invited) (2007)
  • Author(s): K., Inoue
  • Organizer: Gordon Research Conference Apoptotic Cell Recognition & Clearance
  • Place of Presentation: Lewiston, USA
• [Book] ATP receptors in the pain signaling: Clial contribution in neuropathic pain. In Interaction between neurons and glia in aging and disease. (2007)
  • Author(s): Inoue, K
  • Total Pages: 14
  • Publisher: Springer
• [Book] ATP。痛み研究のアプローチ。(河谷正仁編) (2006)
  • Author(s): 井上和秀
  • Total Pages: 10
  • Publisher: 真興交易(株)医書出版部
• [Remarks]
  • URL: http://210.233.60.66/~yakkou/
• [Publications] M.Tsuda, Y.Shigemoto-Mogami, S.Koizumi, A.Mizokoshi, S.Kohsaka, M.W.Salter, K.Inoue: "P2X4 receptors induced in spinal microglia gate tactile allodynia after nerve injury."Nature. 424. 778-783 (2003)
• [Publications] S.Koizumi, K.Fujishita, M.Tsuda, Y.Shigemoto-Mogami, A.Kita, K.Inoue.: "A dynamic regulation by astrocytic ATP of excitatory synaptic transmission cultured rat hippocampal neurons."Pro.Natl.Acad.Sci.USA. 100. 11023-11028 (2003)
• [Publications] T.Moriyama, T.Iida, K.Kobayashi, T.Higashi, T.Fukuoka, H.Tsumura, C.Leon, N.Suzuki, K.Inoue, C.Gachet, K.Noguchi, M.Tominaga: "Possible involvement of P2Y2 metabotropic receptors in ATP-induced transient receptor potential vanilloid receptor 1-mediated thermal hypersensitivity."J.Neurosci.. 23. 6058-6062 (2003)
• [Publications] T.Suzuki, I.Hide, K.Ido, A.Inoue, S.Kohsaka, K.Inoue, Y.Nakata.: "Production and release of neuroprotective TNF by P2X7 receptor-activated microglia."J.Neurosci.. 24. 1-7 (2004)
• [Publications] M.Tsuda, A.Mizokoshi, Y.Shigemoto-Mogami, S.Koizumi, K.Inoue: "Activation of p38MAPK in spinal hyperactive microglia contributes to neuropathic pain hypersensitivity following peripheral nerve injury."Glia. 89. 89-95 (2004)
• [Publications] S.Koizumi, K.Fujishita, K.Inoue, Y.Shigemoto-Mogami, M.Tsuda, K.Inoue.: "Ca2+ waves in keratinocytes are transmitted to sensory neurons : the involvement of extracellular ATP and P2Y2 receptor activation."Biochem.J.. 379. 1-10 (2004)
• [Publications] 井上和秀: "慢性疼痛とミクログリア:ATP受容体の関与.痛みの基礎と臨床"真興交易(株)医書出版部. 93-101 (2003)