{"created":"2023-05-15T15:17:00.347574+00:00","id":3005,"links":{},"metadata":{"_buckets":{"deposit":"127709c3-0b2c-4d82-be73-ccc5c41f0156"},"_deposit":{"created_by":12,"id":"3005","owners":[12],"pid":{"revision_id":0,"type":"depid","value":"3005"},"status":"published"},"_oai":{"id":"oai:obihiro.repo.nii.ac.jp:00003005","sets":["242:243"]},"author_link":["318","178"],"item_6_alternative_title_1":{"attribute_name":"その他（別言語等）の研究課題名","attribute_value_mlt":[{"subitem_alternative_title":"Role of integrin-linked kinaae in neurite outgrowth and axon guidance on neuronal repair","subitem_alternative_title_language":"en"}]},"item_6_contributor_5":{"attribute_name":"研究分担者","attribute_value_mlt":[{"contributorAffiliations":[{"contributorAffiliationNameIdentifiers":[{"contributorAffiliationNameIdentifier":"","contributorAffiliationScheme":"ISNI","contributorAffiliationURI":"http://www.isni.org/isni/"}],"contributorAffiliationNames":[{"contributorAffiliationName":"","contributorAffiliationNameLang":"ja"}]}],"contributorNames":[{"contributorName":"Nishimura, Masakazu","lang":"en"},{"contributorName":"西村, 昌数","lang":"ja"}],"familyNames":[{},{}],"givenNames":[{},{}],"nameIdentifiers":[{"nameIdentifier":"178","nameIdentifierScheme":"WEKO"},{"nameIdentifier":"9000005514215","nameIdentifierScheme":"CiNii ID","nameIdentifierURI":"http://ci.nii.ac.jp/nrid/9000005514215"},{"nameIdentifier":"50011995","nameIdentifierScheme":"e-Rad","nameIdentifierURI":"https://kaken.nii.ac.jp/ja/search/?qm=50011995"},{"nameIdentifier":"read0167489","nameIdentifierScheme":"researchmap","nameIdentifierURI":"https://researchmap.jp/read0167489"}]}]},"item_6_description_10":{"attribute_name":"研究代表者番号","attribute_value_mlt":[{"subitem_description":"50264809","subitem_description_type":"Other"}]},"item_6_description_11":{"attribute_name":"研究機関","attribute_value_mlt":[{"subitem_description":"帯広畜産大学","subitem_description_type":"Other"}]},"item_6_description_13":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"モルモット大脳からクローニングしたIntegrin-linked kinase(ILK)のcDNA(GenBank Accession No.AF256520)をオリゴヌクレオチド変異法により、ILKの触媒ドメインに存在するN末端から359番目のグルタミンサン(E)をリジン(K)に点変異させることで、他の基本ドメインを保存させたまま酵素活性のみを不活化させたドミナントネガティブ体(DN-ILK)を作製した。DN-ILKと野生型ILKは、神経未分化細胞であるN1E-115細胞に導入後、それらタンパク質を高発現させることによりILKの機能を分化条件と未分化条件の両条件下で評価した。N1E-115細胞には、内在ILKが発現しているのでDN-ILKをステイブル発現することにより内在ILKの活性を阻害した結果、N1E-115細胞の神経突起形成を有意に阻害した。それに対して、野生型ILKをトランジエントに高発現させると、非分化条件下にもかかわらず、N1E-115細胞は神経突起を形成し神経へと分化した。以上の結果から、ILKはN1E-115細胞の神経突起形成に必要不可欠であることがわかった。さらに、ILKのシグナル下流にはp38MAP kinaseが存在し、ILKの活性化後に生じるp38MAP kinaseの活性化がN1E-115細胞の神経突起形成に必要であることを証明した。さらに、DN-ILKをステイブル発現したN1E-115細胞のタウタンパク質は高度に異常リン酸化され、その結果、この高度異常リン酸化タウが線維状に形質膜を内面から覆う篭状の形態をとることを新たに発見した。さらにタウの高度異常リン酸化は、内在ILKの不活化に伴い活性化されたGSK-3βにより引き起こされ、内在ILKはGSK-3βのSer9をリン酸化することで不活化し、タウタンパク質が高度異常リン酸化されないように保護していることを証明した。また、タウの異常リン酸化は、DN-ILK発現細胞の神経分化不全に部分的に関与していることを明らかになった。異常リン酸化タウは、アルツハイマー病の病理学的所見で認められる神経原線維変性の発生機構の1つと考えられており大変興味深い。このように、ILKは神経分化機構に直接関与するだけでなく、タウのリン酸化を調節することで、神経分化時の微小管形成の促進と分化後の微小管の安定化に寄与する。","subitem_description_language":"ja","subitem_description_type":"Abstract"},{"subitem_description":"Mouse N1E-115 cells grown on a laminin matrix exhibit neurite outgrowth in response to serum deprivation. Treatment of cells with an antibody against β1 integrin inhibits neurite outgrowth. Thus, β1 integrin is involved in the neuritogenesis of N1E-115 cells on a laminin matrix. Integrin-linked kinase (ILK), a recently identified cytoplasmic serine/threonine protein kinase that binds to the cytoplasmic domain of β1 integrin, has an important role in transmembrane signal transduction via integrins. We report that ILK is expressed in N1E-115 cells, the expression levels of which are constant under both normal and differentiating conditions. A stable transfection of a kinase-deficient mutant of ILK (DN-ILK) results in inhibition of neurite outgrowth in serum-starved N1E-115 cells grown on laminin. On the other hand, a transient expression of wild-type ILK stimulated neurite outgrowth. The ILK activity in the parental cells was transiently activated after seeding on the laminin matrix, whereas that in the DN-ILK transfected cells was not. These results suggest that transient activation of ILK is required for neurite outgrowth in serum-starved N1E-115 cells on laminin. Under the same conditions, p38 mitogen-activated protein (MAP) kinase, but neither MAP kinase/extracellular signal-regulated kinases kinase (MEK) nor extracellular signal-regulated kinases (ERK), was transiently activated after N1E-115 cell attachment to laminin, but not in the DN-ILK expressed cells. The tune course of p38 MAP kinase activation was very similar to that of ILK activation. Furthermore, a p38 MAP kinase inhibitor, SB203580, significantly blocked neurite outgrowth. Thus, activation of p38 MAP kinase is involved in ILK-mediated signal transduction leading to integrin-dependent neurite outgrowth in N1E-115 cells. Moreover, ILK also controls tau phosphorylation via regulation of glycogen synthase kinase-3β (GSK-3β) activity in N1E-115 cells. Stable transfection of DN-ILK resulted in aberrant tau phosphorylation in N1E-115 cells at sites recognized by the Tau-1 antibody, which are identical to some of the phosphorylation sites in paired helical filaments, (PHF)-tau, in brains of patients with Alzheimer's disease. The tau phosphorylation levels in the DN-ILK-expressing cells are constant under normal and differentiating conditions. On the other hand, aberrant tau phosphorylation was not observed in the parental control cells. ILK inactivation resulted in an increase in the active form but a decrease in the inactive form of GSK-3β, which is a candidate kinase involved in PHF-tau formation. Moreover, inhibition of GSK-3β with lithium prevented aberrant tau phosphorylation in the DN-ILK-expressing cells. These results suggest that ILK inactivation results in aberrant tau phosphorylation via sustained activation of GSK-3β in N1E-115 Cells. ILK directly phosphorylates GSK-3β and inhibits its activity. Therefore, endogenous ILK protects against GSK-3β-induced aberrant tau phosphorylation via inhibition of GSK-3β activity in N1E-115 cells.","subitem_description_language":"en","subitem_description_type":"Abstract"}]},"item_6_description_14":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"平成14年度～平成15年度科学研究費補助金基盤研究(C)(2)研究成果報告書","subitem_description_language":"ja","subitem_description_type":"Other"}]},"item_6_description_19":{"attribute_name":"フォーマット","attribute_value_mlt":[{"subitem_description":"application/pdf","subitem_description_type":"Other"}]},"item_6_description_9":{"attribute_name":"研究課題番号","attribute_value_mlt":[{"subitem_description":"14560242","subitem_description_type":"Other"}]},"item_6_text_12":{"attribute_name":"助成元","attribute_value_mlt":[{"subitem_text_language":"ja","subitem_text_value":"科学研究費助成事業"}]},"item_6_version_type_20":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_970fb48d4fbd8a85","subitem_version_type":"VoR"}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2018-01-30"}],"displaytype":"detail","filename":"ILK.pdf","filesize":[{"value":"1.6 MB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"ILK.pdf","url":"https://obihiro.repo.nii.ac.jp/record/3005/files/ILK.pdf"},"version_id":"722290c3-29b7-4b63-b4ec-2f8ed51c3c13"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"jpn"}]},"item_researcher":{"attribute_name":"研究代表者","attribute_type":"creator","attribute_value_mlt":[{"creatorAffiliations":[{"affiliationNameIdentifiers":[{"affiliationNameIdentifier":"","affiliationNameIdentifierScheme":"ISNI","affiliationNameIdentifierURI":"http://www.isni.org/isni/"}],"affiliationNames":[{"affiliationName":"","affiliationNameLang":"ja"}]}],"creatorNames":[{"creatorName":"Ishii, Toshiaki","creatorNameLang":"en"},{"creatorName":"石井, 利明","creatorNameLang":"ja"}],"familyNames":[{},{}],"givenNames":[{},{}],"nameIdentifiers":[{},{},{},{}]}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"research report","resourceuri":"http://purl.org/coar/resource_type/c_18ws"}]},"item_title":"神経修復時の神経突起、軸索ガイダンスに及ぼすインテグリンリンクドキナーゼの役割","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"神経修復時の神経突起、軸索ガイダンスに及ぼすインテグリンリンクドキナーゼの役割","subitem_title_language":"ja"}]},"item_type_id":"6","owner":"12","path":["243"],"pubdate":{"attribute_name":"PubDate","attribute_value":"2010-10-20"},"publish_date":"2010-10-20","publish_status":"0","recid":"3005","relation_version_is_last":true,"title":["神経修復時の神経突起、軸索ガイダンスに及ぼすインテグリンリンクドキナーゼの役割"],"weko_creator_id":"12","weko_shared_id":-1},"updated":"2024-05-28T02:24:56.182449+00:00"}