2000 年度実績報告書

DNAマイクロアレイを利用したてんかん治療薬の新規標的分子の検索

研究課題

研究課題/領域番号	12877016
研究機関	神戸大学
研究代表者	斎藤尚亮神戸大学, バイオシグナル研究センター, 教授 (60178499)
研究分担者	酒井規雄神戸大学, バイオシグナル研究センター, 助教授 (70263407)
キーワード	てんかん / グルタミン酸トランスポーター / ノックアウトマウス / DNAマイクロアレイ
研究概要	DNAマイクロアレイを用いたてんかん関連分子の検索グルタミン酸トランスポーター(GLT-1)ノックアウト(KO)マウス脳と野生型マウス脳からそれぞれmRNAを抽出し、それらを鋳型にしてランダムプライマーを用いてcDNAプローブを作製し、DNAマイクロアレイに適用した。GLT-1KOマウスと野生型マウスの遺伝子発現のプロフィールの差異をコンピューター解析し、GLT-1KOマウスにおいて特異的に発現が増加あるいは減少している遺伝子を検索した。その結果、複数の候補遺伝子を同定した。候補遺伝子の中で注目されるものとして、神経特異的機能タンパク質、特に神経伝達物質分泌機構に関連するタンパク質が挙げられる。今後、この候補遺伝子に特に注目し、1)ヘテロ型GLT-1KOマウスにおいても、発現に差異がでるものを選出する、2)電気ショックなどで人為的に痙攣発作を誘発したマウスと正常マウスを用いてDNAマイクロアレイを行い、発現に差異が出た遺伝子と1)で発現に差異が出た遺伝子を比較し、一致するものは、てんかん発作自身によって発現が変化した遺伝子と判断し、本研究の目的遺伝子から除外する、などを行い、てんかんの発症や予防に関連し、さらに、新しい抗てんかん薬の標的になりうる分子を明らかにしてゆく。

研究成果

(15件)

すべてその他

すべて文献書誌 (15件)

[文献書誌] Kajimoto,T.: "Subtype-specific translocation of δ-PKC and its activation by tyrosine phosphorylation induced by ceramide in HeLa cells."Mol.Cell.Biol.. (印刷中). (2001)
[文献書誌] Ohmori,S.: "Importance of PKC targeting for the phosphorylation of its substrate. myristoylated alanine-rich C-kinase substrate (MARCKS)."J.Biol.Chem.. 275. 26449-26457 (2000)
[文献書誌] Shirai,Y.: "Subtype-specific translocation of diacylglycerol kinase α and γ and its correlation with protein kinase C."J.Biol.Chem.. 275. 24760-24766 (2000)
[文献書誌] Larsen,E.C.: "Differential requirement for classic and novel PKC isoforms in respiratory hurst and phagocytosis in RAW 264.7 cells."J.Immunol.165. 2809-2817 (2000)
[文献書誌] Shirai,Y.: "Phospholipase A2 and its products are involved in the purinergic rcceptormediated translocation of protein kinase C in CHO-K1 cells."J.Cell Sci.. 113. 1335-1343 (2000)
[文献書誌] Sumioka,K.: "Induction of 55 kDa PKN cleavage product by ischemia/reperfusion model in the rat retina."Invest.Opthal.Vis.Sci.,. 41. 29-35 (2000)
[文献書誌] Kondoh,T.: "In vivo gene transfer into periventricular region by electroporation."Neurologia medico-chirurgica.. 40. 618-623 (2000)
[文献書誌] Sakai,N.: "Involvement of the actin cytoskeleton in the regulation of serotonin transporter (SET) activity : possible mechanism underlying SET regulation by protein kinase C."Neurochem.Int.. 36. 567-579 (2000)
[文献書誌] Oyasu,M.: "Immunocytochemical localization of a neuron-specific thrombospondin-1-like protein, NELL2 : Light and electron microscopic studies in the rat brain."Mol.Brain Res.. 76. 151-160 (2000)
[文献書誌] Yokogawa,T.: "Evidence that 3'-phosphorylated polyphosphoinositides are generated at the nuclear surface : use of immunostaining technique with monoclonal antibodies specific for PI3,4-P2"FEBS Lett.. 473. 222-226 (2000)
[文献書誌] Akiyama,H.: "Blood-brain barrier formation of grafted human umbilical vein endothelial cells in athymic mouse brain."Brain Res.. 858. 172-176 (2000)
[文献書誌] Ueyama,T.: "cDNA cloning of an alternative splicing variant of protein kinase Cδ (PKCδIII), a new truncated form of PKCδ, in rats."Biochem.Biophys.Res.Commun.. 269. 557-563 (2000)
[文献書誌] Nakagawa,N., 他: "ENG, containing PDZ and LIM domain, heart/skeletal muscle-specific protein, associates with cytoskeletal protein through its PDZ domain."Biochem.Biophys.Res.Commum.. 272. 505-512 (2000)
[文献書誌] Shibaguchi,H., 他: "Role of synaptophysin in exocytic release of dopamine from Xenopus oocytes injected with rat brain mRNA."Cellular and Molecular Neurobiology.. 20. 401-408 (2000)
[文献書誌] Nakai,M., 他: "Amyloidβ protein activates PKC-δ and induces translocation of myristoylated alanine-rich C kinase substrate (MARCKS) in microglia."Neurochem.Int.. (印刷中).

2000 年度 実績報告書

DNAマイクロアレイを利用したてんかん治療薬の新規標的分子の検索

研究代表者

斎藤 尚亮 神戸大学, バイオシグナル研究センター, 教授 (60178499)

研究成果

[文献書誌] Kajimoto,T.: "Subtype-specific translocation of δ-PKC and its activation by tyrosine phosphorylation induced by ceramide in HeLa cells."Mol.Cell.Biol.. (印刷中). (2001)

[文献書誌] Ohmori,S.: "Importance of PKC targeting for the phosphorylation of its substrate. myristoylated alanine-rich C-kinase substrate (MARCKS)."J.Biol.Chem.. 275. 26449-26457 (2000)

[文献書誌] Shirai,Y.: "Subtype-specific translocation of diacylglycerol kinase α and γ and its correlation with protein kinase C."J.Biol.Chem.. 275. 24760-24766 (2000)

[文献書誌] Larsen,E.C.: "Differential requirement for classic and novel PKC isoforms in respiratory hurst and phagocytosis in RAW 264.7 cells."J.Immunol.165. 2809-2817 (2000)

[文献書誌] Shirai,Y.: "Phospholipase A2 and its products are involved in the purinergic rcceptormediated translocation of protein kinase C in CHO-K1 cells."J.Cell Sci.. 113. 1335-1343 (2000)

[文献書誌] Sumioka,K.: "Induction of 55 kDa PKN cleavage product by ischemia/reperfusion model in the rat retina."Invest.Opthal.Vis.Sci.,. 41. 29-35 (2000)

[文献書誌] Kondoh,T.: "In vivo gene transfer into periventricular region by electroporation."Neurologia medico-chirurgica.. 40. 618-623 (2000)

[文献書誌] Sakai,N.: "Involvement of the actin cytoskeleton in the regulation of serotonin transporter (SET) activity : possible mechanism underlying SET regulation by protein kinase C."Neurochem.Int.. 36. 567-579 (2000)

[文献書誌] Oyasu,M.: "Immunocytochemical localization of a neuron-specific thrombospondin-1-like protein, NELL2 : Light and electron microscopic studies in the rat brain."Mol.Brain Res.. 76. 151-160 (2000)

[文献書誌] Yokogawa,T.: "Evidence that 3'-phosphorylated polyphosphoinositides are generated at the nuclear surface : use of immunostaining technique with monoclonal antibodies specific for PI3,4-P2"FEBS Lett.. 473. 222-226 (2000)

[文献書誌] Akiyama,H.: "Blood-brain barrier formation of grafted human umbilical vein endothelial cells in athymic mouse brain."Brain Res.. 858. 172-176 (2000)

[文献書誌] Ueyama,T.: "cDNA cloning of an alternative splicing variant of protein kinase Cδ (PKCδIII), a new truncated form of PKCδ, in rats."Biochem.Biophys.Res.Commun.. 269. 557-563 (2000)

[文献書誌] Nakagawa,N., 他: "ENG, containing PDZ and LIM domain, heart/skeletal muscle-specific protein, associates with cytoskeletal protein through its PDZ domain."Biochem.Biophys.Res.Commum.. 272. 505-512 (2000)

[文献書誌] Shibaguchi,H., 他: "Role of synaptophysin in exocytic release of dopamine from Xenopus oocytes injected with rat brain mRNA."Cellular and Molecular Neurobiology.. 20. 401-408 (2000)

[文献書誌] Nakai,M., 他: "Amyloidβ protein activates PKC-δ and induces translocation of myristoylated alanine-rich C kinase substrate (MARCKS) in microglia."Neurochem.Int.. (印刷中).

2000 年度実績報告書

斎藤尚亮神戸大学, バイオシグナル研究センター, 教授 (60178499)