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トランスグルタミナーゼの神経系における役割りについて: 神経再生の分子機序解明
https://doi.org/10.24517/00034807
https://doi.org/10.24517/000348076b55a080-7351-4fe6-ae0e-626563f66f01
名前 / ファイル | ライセンス | アクション |
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ME-PR-KATO-S-kanen 2002-7p.pdf (242.2 kB)
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Item type | 報告書 / Research Paper(1) | |||||
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公開日 | 2017-10-05 | |||||
タイトル | ||||||
タイトル | トランスグルタミナーゼの神経系における役割りについて: 神経再生の分子機序解明 | |||||
タイトル | ||||||
言語 | en | |||||
タイトル | A role ofIransglutaminase during optic nerve regeneration in the goldfish | |||||
言語 | ||||||
言語 | jpn | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_18ws | |||||
資源タイプ | research report | |||||
ID登録 | ||||||
ID登録 | 10.24517/00034807 | |||||
ID登録タイプ | JaLC | |||||
著者 |
加藤, 聖
× 加藤, 聖 |
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著者別表示 |
Kato, Satoru
× Kato, Satoru |
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書誌情報 |
平成13(2001)年度 科学研究費補助金 基盤研究(C) 研究成果報告書 en : 2001 Fiscal Year Final Research Report 巻 2000-2001, p. 7p., 発行日 2002-03-01 |
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出版者 | ||||||
出版者 | 金沢大学大学院医学系研究科 | |||||
抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | 金魚の視神経は哺乳類と異なり再生し、4〜6ヵ月で視覚機能も完全に回復する。この長い再生過程(4〜6ヵ月)で働く分子を同定しその機能を追求することはきわめて重要である。本研究では特に視神経切断後早期に切断端からの再生神経線維の伸長に働く分子の同定を目指した。視神経切断後5日目の網膜からmRNAを抽出しcDNAを合成しcDNAライブラリーを作製した。ディファレンシャルハイブリ法で対照(無処置)と比べて視神経切断5日目に発現が増加(upregulation)するcDNAクローンをスクリーニングした。そのクローンの塩基配列を決定しDNA data baseの検索から、トランスグルタミネース(TG)やNa, K-ATPase α 3サブユニットをコードする遺伝子が捕捉された。ISH法や免疫組織化学法によりTGやNa, K-ATPase α 3の視神経切断後の経時変化を調べた所、網膜神経節細胞にのみ切断早期に発現が上昇し、Na, K-ATPase α 3は10日目、TGは20日でピークとなりその後徐々に対照群のそれに復した。また、成熟金魚の視神経をあらかじめ切っておき(5〜7日前)、網膜片を作製し培養すると神経節細胞から神経突起が伸長(神経再生のin vitro系の確立)する。この培養系にTG抗体やNa, K-ATPase α 3の特異的阻害剤ウアバインを添加すると、神経突起の伸長が完全に抑えられた。このことからTGやNa, K-ATPase α 3は視神経再生初期の再生線維の伸長に重要な役割を果たしていることがわかる。今後この遺伝子クローニング法を駆使して再生各時期に働く遺伝子を見つけたいと思っている。また、遺伝子改変動物を作製し、その遺伝子の機能も明らかにしたい。 | |||||
抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | The goldfish optic nerve can regenerate after injury. Regrowth of optic nerve fibers starts to extend 7-10 days after optic nerve transection. They arrive at the optic tectum 3-5 weeks after axtfomy. Finally, visual function is restored 4-6 months after optic nerve transection. To understand the molecular mechanism of optic nerve regeneration, we identified genes whose expression is specifically upregulated during the early stage of optic nerve regeneration. A cDNA library constructed from goldfish retina 5 days after transection was screened Two clones were identified as the Na, K-ATPase catalytic subunitalpha 3 isoform and the neural type of transglutaminase (TG) by sequence analysis. In northern hybridization, the expression level of the both mRNAs was significantly increased at 2 days and peaked at 5-10 days, and then gradually decreased and returned to control level by 45-60 days after optic nerve transection. In situ hybridization have revealed the location of this transient retinal change after axotomy. The increased expression was observed only in the ganglion cell layer at 10-20 days after optic nerve transection, The cDNA of TG was inserted into the plasmid DNA and thus a fused protein of TG (ca. 70kDa) was obtained. Antiserum against the recombinant TG was made by immunization. Immunohistochemical staining further confirmed the data of in situ hybridization. In an explant culture system, neurite outgrowth fkm the retina 7 days after optic nerve transection was spontaneously promoted. A low concentration of ouabain (an inhibitor of Na,K ATPase alpha 3 subunit) or neutral antibody against TG, completely blocked the spontaneous neurite outgrowth from the lesioned retina. Together, these data indicate that upregulation of the Na, K-ATPase alpha 3 subunit or TG is involved in the regrowth of ganglion cell axons after axotomy. Such molecular techniques will become useful tool for future genetic studies of nerve regeneration. | |||||
内容記述 | ||||||
内容記述タイプ | Other | |||||
内容記述 | 研究課題/領域番号:12680750, 研究期間(年度):2000–2001 | |||||
内容記述 | ||||||
内容記述タイプ | Other | |||||
内容記述 | 出典:「トランスグルタミナーゼの神経系における役割りについて: 神経再生の分子機序解明」研究成果報告書 課題番号12680750 (KAKEN:科学研究費助成事業データベース(国立情報学研究所)) 本文データは著者版報告書より作成 |
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著者版フラグ | ||||||
出版タイプ | AM | |||||
出版タイプResource | http://purl.org/coar/version/c_ab4af688f83e57aa | |||||
関連URI | ||||||
識別子タイプ | URI | |||||
関連識別子 | https://kaken.nii.ac.jp/search/?qm=10019614 | |||||
関連URI | ||||||
識別子タイプ | URI | |||||
関連識別子 | https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12680750/ | |||||
関連URI | ||||||
識別子タイプ | URI | |||||
関連識別子 | https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-12680750/126807502001kenkyu_seika_hokoku_gaiyo/ |