2000 Fiscal Year Annual Research Report

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

Research Project

Project/Area Number	12877016
Research Institution	Kobe University
Principal Investigator	斎藤尚亮神戸大学, バイオシグナル研究センター, 教授 (60178499)
Co-Investigator(Kenkyū-buntansha)	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)
Keywords	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
Research Abstract	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。

Research Products

(15 results)

All Other

All Publications (15 results)

[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] Kondoh,T.: "In vivo gene transfer into periventricular region by electroporation."Neurologia medico-chirurgica.. 40. 618-623 (2000)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] Akiyama,H.: "Blood-brain barrier formation of grafted human umbilical vein endothelial cells in athymic mouse brain."Brain Res.. 858. 172-176 (2000)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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 Fiscal Year Annual Research Report

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

Principal Investigator

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

Research Products

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

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

[Publications] 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)

[Publications] 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)

[Publications] 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)

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

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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).

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