ミクログリアによるグルタミン酸を介した神経伝達の動的制御

• Project/Area Number: 15082204
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Kyushu University
• Principal Investigator: 中西 博 Kyushu University, 大学院・歯学研究院, 教授 (20155774)
• Co-Investigator(Kenkyū-buntansha): 武 洲 九州大学, 大学院・歯学研究院, 助教 (10420598) ; 田中 嘉孝 九州大学, 大学院・薬学研究院, 教授 (20217095) ; 門司 晃 九州大学, 医学部附属病院, 助手 (00294942)
• Project Period (FY): 2003 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥61,400,000 (Direct Cost: ¥61,400,000); Fiscal Year 2007: ¥11,500,000 (Direct Cost: ¥11,500,000); Fiscal Year 2006: ¥11,500,000 (Direct Cost: ¥11,500,000); Fiscal Year 2005: ¥12,800,000 (Direct Cost: ¥12,800,000); Fiscal Year 2004: ¥12,800,000 (Direct Cost: ¥12,800,000); Fiscal Year 2003: ¥12,800,000 (Direct Cost: ¥12,800,000)
• Keywords: ミクログリア / グルタミン酸 / 神経伝達 / synaptic stripping / ATP / 培養上清 / dendritic beading / パッチクランプ / カイニン酸受容体 / 細胞周期 / リアルタイムRT-PCR / GluR / 脳虚血 / 海馬CA1野ニューロン / NMDA受容体 / グリシン結合部位 / HPLC / グリシン / パッチクランプ法 / 長期増強

Research Abstract

交付申請書の研究実施計画に沿って今年度は以下の2点についての解析を行った。
(1)Synaptic strippingによりミクログリアがグルタミン酸を介した神経伝達を遮断する可能性の検討
ミクログリアは偽足をシナプス間隙に差し込むことによりシナプス終末部を軸索損傷ニューロの表面から引き離し(synaptic stripping)、神経伝達を遮断することが示唆されているが不明な点も多い。ラット迷走神経背側運動(DMV)ニューロンの興奮性シナプス電流頻度は軸索切断2日目より有意に減少した。シナプス電流頻度の減少はミクログリアに取り囲まれていないDMVニューロンでも認められた。また、減少したシナプス電流頻度は高K^+刺激ならびにP2受容体拮抗剤PPADSは減少したシナプス電流頻度を回復させた。一方、機械的に単離したラットDMVニューロンでは軸索切断に伴うシナプス電流頻度の減少は認められなかった。以上の結果より、軸索切断に伴って主にグリア細胞から放出されたATPがグルタミン酸を介した神経伝達を抑制し、ミクログリアの神経伝達への影響は少ないことが示唆された。
(2)ミクログリアの産生する液性因子により誘発させるdendritic beading形成抑制作用
疎水カラムを用いて分画したサンプルではカラムに吸着する分画に活性が認められた。一方、陽イオン交換樹脂カラムを用いて分画したサンプルではカラムに吸着が弱い分画に活性が認められた。疎水カラム及び陽イオン交換樹脂カラムを用いて分画したサンプルに関する実験においてミクログリア培養上清の分画サンプルにNMDAによって誘発されるdendritic beading形成に対する抑制活性が認められており、培養液のみの分画サンプルには活性は全く認められなかった。このことから活性分子は疎水性であることが示唆された。

Research Products

• [Journal Article] Involvement of PGE_2 released from leptomeningeal cells in increased expression of TGF-β1 in glial cells and cortical neurons during systemic inflammation. (2007)
  • Author(s): Wu, Z., et. al.
  • Journal Title: Journal of Neuroscience Research 85 Pages: 184-192
  • Peer Reviewed
• [Journal Article] Phospholipids modulate superoxide and nitric oxide production by lipoplysaccharide and phorbol 12-myristate-13-acetate-activated microglia. (2007)
  • Author(s): Hashioka, S., et. al.
  • Journal Title: Neurochemistry International 50 Pages: 499-506
  • Peer Reviewed
• [Journal Article] Phospholipids modulate superoxide and nitric oxide production by lipoplysaccharide and phorbol 12-myristate-13-acetate-activated microglia. (2007)
  • Author(s): Hashioka, S., et. al.
  • Journal Title: Free Radical Biology and Medicine 42 Pages: 945-954
  • Peer Reviewed
• [Journal Article] Impairment of the microglial responses to the facial berve axotomy in cathepsin S-deficient mice. (2007)
  • Author(s): Hao, H. P., et. al.
  • Journal Title: Journal of Neuroscience Research 85 Pages: 2196-2206
  • Peer Reviewed
• [Journal Article] Involvement of lysosomal storage-induced p38 MAP kinase activation in the overproducection of nitric oxide by microglia in cathepsin D-deficient mice. (2007)
  • Author(s): Yamasaki, R., et. al.
  • Journal Title: Molecular and Cellular Neuroscience 35 Pages: 573-584
  • Peer Reviewed
• [Journal Article] Involvement of PGE_2 released from leptomeningeal cells in increased expression of TGF-β1 in glial cells and cortical neurons during systemic unflammation. (2007)
  • Author(s): Wu, Z.et al.
  • Journal Title: J. Neurosci. Res. 85・1 Pages: 184-192
• [Journal Article] Phospholipids modulate superoxide and nitric oxide production by lipopolysaccharide and phorbol 12-myristate-13-acetate-activated microglia. (2007)
  • Author(s): Hashioka, S. et al.
  • Journal Title: Neurochem. Int. 50・3 Pages: 499-506
• [Journal Article] Phosphatidylserine and phosphatidylcholine-containing liposomes inhibit amyloid β and interferon-γ-induced microglial activation. (2007)
  • Author(s): Hashioka, S. et al.
  • Journal Title: Free Radic. Biol. Med. 42・7 Pages: 945-954
• [Journal Article] The intra-arterial injection of microglia protects hippocampal CAI neurns against global ischemia-induced functional deficits in rats. (2006)
  • Author(s): Hayashi, Y. et al.
  • Journal Title: Neuroscience 142・1 Pages: 87-96
• [Journal Article] Cell cycle-dependent regulation of kainite-induced inward currents in microglia. (2006)
  • Author(s): Yamada, J. et al.
  • Journal Title: Biochem. Biophys. Res. Commun. 349・3 Pages: 913-919
• [Journal Article] Involvement of COX-1 and up-regulated prostaglandin E synthases in phosphatidylserine liposome-induced prostaglandin E_2 production of microglia. (2006)
  • Author(s): Zhang, J. et al.
  • Journal Title: J.Neuroimmunol. 172・1-2 Pages: 112-120
• [Journal Article] Potentiation of the NMDA receptor-mediated responses through the activation of the glycine site by microglial secreting soluble factor. (2006)
  • Author(s): Hayashi, Y. et al.
  • Journal Title: Glia 53・6 Pages: 660-668
• [Journal Article] Amyloid-β fibril formation is not necessarily required for microglial activation by the peptides (2005)
  • Author(s): Hashioka, S. et al.
  • Journal Title: Neurochem.Int. 47・5 Pages: 369-376
• [Journal Article] Proteolytic degradation of glutamate decarboxylase mediates disinhibition of hippocampal CA3 pyramidal calls in cathepsin D-deficient mice. (2005)
  • Author(s): Shimizu, T. et al.
  • Journal Title: J.Neurochem. et al. 94・3 Pages: 680-690
• [Journal Article] Leptomeningeal cells activate microglial and astrocytes to induce IL-10 production by releasing pro-inflammatory cytokines during systemic inflammation. (2005)
  • Author(s): Wu, Z.et al.
  • Journal Title: J.Neuroimmunol. 167・1-2 Pages: 90-98
• [Journal Article] Sibutramine induces potential-dependent excitosis release but not carrier-mediated release of dopamine and 5-hydroxytryptamine (2004)
  • Author(s): Ukai, K. et al.
  • Journal Title: Eur.J.Pharmacol. 484 Pages: 209-215
• [Journal Article] Amyloid-β fibril formation is not necessarily required for microglial activation by the peptidee.
  • Author(s): Hasioka, S. et al.
  • Journal Title: Neurochem Int. (in press)
• [Presentation] Synaptic Strippingにおける細胞外ヌクレオチドの関与 (2008)
  • Author(s): 中西 博
  • Organizer: 第81回日本薬理学会総会、シンポジウム:細胞外ヌクレオチドによる脳機能制御-分子ダイナミクスと疾病への関与
  • Place of Presentation: 横浜
  • Year and Date: 2008-03-17
• [Presentation] Modulaton of NMDA receptor-mediated responses by microglial secreting soluble factors. (2007)
  • Author(s): Hayashi, Y・Nakanishi, H.
  • Organizer: 第37回北米神経科学会
  • Place of Presentation: サンディエゴ
  • Year and Date: 2007-11-07
• [Presentation] Systemic inflammation induces age-dependent defferential activation of gilal cell sthrough the leptomeninges. (2007)
  • Author(s): Wu, Z・Nakanishi, H.
  • Organizer: 第8回ヨーロッパグリア(Euroglia) 会議
  • Place of Presentation: ロンドン
  • Year and Date: 2007-09-06
• [Book] Age-dependent differential activation profiles of glial cells by the leptomeninges during systemic inflammation: In Neuroimmmunology Research Focus,(Broglio PV ed) (2007)
  • Author(s): Wu, Z・Nakanishi, H.(分担)
  • Publisher: Nova Science Publishers Inc.
• [Book] Microglial Proteases: Ineural Protein Metabolism and Function. Handbook of Neurochemistry and Molecular Neurobiology, Vol.7(LajthaA ed) (2007)
  • Author(s): Nakanishi, H.(分担)
  • Publisher: Kluwer Publishing Corporation
• [Book] Proteases in the Brain(Proteases and synaptic activity) (2005)
  • Author(s): Nakanishi, H.
  • Total Pages: 383
  • Publisher: Kluwer Academic/Plenum Publisher
• [Book] Protease in Biology and Disease, Chapter 12. Protease and Synaptic Activity pp. 303-〜322 (2005)
  • Author(s): Nakanishi, H.
  • Total Pages: 383
  • Publisher: Kluwer Academic/Plenum Publisher
• [Publications] Moriguchi, S.et al.: "Potentiation of NMDA receptor-mediated synaptic responses by microglia."Molecular Brain Research. 119. 160-169 (2003)
• [Publications] Okada, M.et al.: "Pepstatin A induces extracellular acidification distinct from aspartic protease inhibition in microglial cell line"Glia. 43. 167-174 (2003)
• [Publications] Zhang, J.et al.: "Activation of m-calpain in the developing cortical neurons following Methylmercury treatment."Developmental Brain Research. 142. 105-110 (2003)
• [Publications] Okada, M.et al.: "How dose prolonged caloric restriction ameliorate age-related impairment Of long-term potentiation in the hippocampus?"Molecular Brain Research. 111. 175-181 (2003)
• [Publications] Koike, M.et al.: "The involvement of caspase-dependent and -independent neuronal death in retinal atrophy of cathepsin D-deficient mice."Molecular Cellular Neuroscience. 22. 146-161 (2003)
• [Publications] Nakanishi, H.: "Neuronal and microglial cathepsins in ageing and age-related diseases."Ageing Research Review. 2. 367-381 (2003)
• [Publications] Nakanishi, H.: "Microglial functions and proteases."Molecular Neurobiology. 27. 163-176 (2003)
• [Publications] Nakanishi, H.: "Microglial proteases : strategic targets for neuroprotective agents."Current Neuropharmacology. 1. 99-108 (2003)
• [Publications] 中西 博: "ミクログリアとリソソーム病"脳21. 6. 15-19 (2003)