神経細胞死のメカニズムと防御に関する研究

• 研究課題/領域番号: 03263105
• 研究種目: 重点領域研究
• 配分区分: 補助金
• 研究機関: 順天堂大学
• 研究代表者: 水野 美邦 順天堂大学, 医学部・神経学講座, 教授 (30049043)
• 研究分担者: 桐野 高明 帝京大学, 医学部・脳神経外科, 助教授 (10161605) ; 中野 稔 浜松医科大学, 光量子医学実験センター, 教授 (00008236) ; 吉田 充男 自治医科大学, 神経内科, 教授 (70048966) ; 宝来 聡 国立遺伝学研究所, 総合遺伝研究系, 助教授 (40126157) ; 田中 雅嗣 名古屋大学, 医学部第二生化学, 助教授 (60155166)
• 研究期間 (年度): 1991 – 1992
• 研究課題ステータス: 完了 (1991年度)
• 配分額: 81,000千円 (直接経費: 81,000千円); 1991年度: 81,000千円 (直接経費: 81,000千円)
• キーワード: 神経細胞死 / パ-キンソン病 / ミトコンドリア脳筋症 / ニュ-ロトキシン / フリ-ラジカル / 興奮性アミノ酸 / 脳虚血 / 分子生物学

研究概要

1.エネルギ-産生障害による細胞死:パ-キンソン病黒質神経細胞に複合体Iの低下のあることを免疫組織化学で証明,更に筋にも複合体I活性低下があるが,血小板,リンパ球など寿命の短い細胞では変化していないことを示した.5例のパ-キンソン病患者にてmtDNA全塩基配列を決定,多くの塩基置換を発見.これらの変異により,電子伝達系よりラジカル漏出が増加,mtDNAの酸化的損傷を惹起,点変異,欠失を誘導する可能性を示した.
MELASではmtDNAのtRNA^<Leu(UUR)>遺伝子(塩基番号3243)の点突然変異を40名中32名に発見,残る8名中3名でこれと異なる塩基番号3271のTからCへの点突然変異を発見.更にミスマッチプライマ-法を用いたPCR法により簡便なDNA診断法を開発した.
2.内因性・外因性毒性物質による神経細胞死:1MeTIQが脳内在性物質として存在,MPTP毒性に拮抗すること,Parkinson病で減少していることを明らかにした.一方TIQは黒質神経細胞障害を起こすことを示した.
3.フリ-ラジカルによる細胞障害:MCLA試薬を用い,虚血障害の際浸潤する白血球の出す化学発光をin vivoで捉え,間接的に好中球を算出する方法を開発,又組織片に対する高感度定量法を開発.又培養細胞を用いFe‐ADPとメラニンがsuperoxide anionを介し脂質過酸化を起こす事を示す.
4.興奮性アミノ酸及び虚血による細胞障害:カイニン酸誘導体を合成,既知の興奮性アミノ酸を凌ぐ強力なグルタミン酸アゴニストを発見.又海馬遅発性神経細胞死で細胞内カルシウムイオンが高いレベルで動揺していることを明らかにし,短時間の脳虚血は神経細胞に虚血耐性を誘導,同時にストレス蛋白が発現すること,耐性の発現した細胞では蛋白代謝回転が良好であることを証明した.

研究成果

• [文献書誌] Hattori, N., Tanaka, M., Ozawa, T. and Mizuno,Y.: "Immunohistochemical studies on Complexes I,II,III and IV of mitochondria in Parkinson's disease." Ann. Neurol.30. 563-571 (1991)
• [文献書誌] Hattori, Y., Yoshino, H., Kondo, T., Mizuno, Y. and Horai, S.: "Is Parkinson's disease a“mitochondrial disease?"." J. Neurol. Sci.(1992)
• [文献書誌] Yoshino, H., Nakagawa‐Hattori, Y., Kondo, T. and Mizuno, Y.: "Mitochondrial complex I and II activities of lymphocytes and platelets in Parkinson's disease." J. Neurol. Transm. PD Section. 4. 27-34 (1992)
• [文献書誌] Tanaka, M., et al.: "Mitocondrial DNA mutations in mitochondrial myopathy,encephalopathy,lactic acidosis,and stroke‐like episodes(MELAS)." Biochem. Biophys. Res. Commun.174. 861-868 (1991)
• [文献書誌] Ito, H., et al.: "Mitochondrial leucine tRNA mutation in a mitochondrial encephalomyopathy." Lancet. 337. 234-235 (1991)
• [文献書誌] Ozawa, T., et al.: "Distinct clustering of point mutations in mitochondrial DNA among patients with mitochondrial encephalomyopathies and with Parkinson's disease." Biochem. Biophys. Res. Commun.177. 518-525 (1991)
• [文献書誌] Hattori, K., et al.: "Age‐dependent increase in deleted mitocondrial DNA in the human heart:possible contributory factor to presbycardia." Am. Heart J.121. 1735-1742 (1991)
• [文献書誌] Hayakawa, M., et al.: "Massive combersion of guanosine to 8‐hydroxy‐guanosine in mouse liver mitochondrial DNA by administration of azidothymidine." Biochem. Biophys. Res. Commun.176. 87-93 (1991)
• [文献書誌] Hayakawa, M., et al.: "Age‐associated accumulation of 8‐hydroxydeoxyguanosine in mitochondrial DNA of human diaphragm." Biochem. Biophys. Res. Commun.179. 1023-1029 (1991)
• [文献書誌] Ohno, K., et al.: "Direct DNA sequencing from colony:analysis of multiple deletions of mitochondrial genome." Biochim. Biophys. Acta. 1090. 9-16 (1991)
• [文献書誌] Nakamura, S., et al.: "The histochemical demonstration of monoamine oxidase‐containing neurons in the human hypothalamus." Neuroscience. 44. 457-463 (1991)
• [文献書誌] Kato, H., Liu, Y., Araki, T. and Kogure, K.: "Temporal profile of the effects of pretreatment with brief cerebral ischemia on the neuronal damage following secondary ischemic insult in the gerbil:cumulative damage and protective effects." Brain Res.553. 238-242 (1991)
• [文献書誌] Nakata, N., Kato, H., Liu, Y. and Kogure, K.: "Effects of pretreatment with sublethal ischemia on the extracellular glutamate concentrations during secondary ischemia in the gerbil hippocampus evaluated with intracranial microdialysis." Neurosci. Lett.
• [文献書誌] Kato, H., Araki, T., Hara, H. and Kogure, K.: "Sequential changes in muscarinic acetylcholine,adenosine Al and calcium antagonist binding sites in the gerbil hippocampus following repeated brief ischemia." Brain Res.553. 33-38 (1991)
• [文献書誌] Kato, H., Araki, T., Hara, H. and Kogure, K.: "Alterations in second‐messenger systems in the gerbil hippocampus following repeated brief ischemia:autoradiographic analysis using[^3H] inositol triphosphate,[^3H]phorbol dibutyrate and [^3H]forskolin." Brain Res. Bull.27. 759-765 (1991)
• [文献書誌] Kato, H., Liu, Y., Araki, T. and Kogure, K.: "MK‐801,but not anisomycin,ingibits the induction of tolerance to ischemia in the gerbil hippocampus." Neurosci. Lett.
• [文献書誌] Ono, H., Hasebe, Y., Satoh, M., Nagao, T., Ohta, S., Hirobe, M. and Fukuda, H.: "Amphetamine‐antagonistic property of 4‐pheny1‐1,2,3,4‐tetrahydroisoquinoline:inhibition of spinal reflex‐enhancing effects of methamphetamine,phenylethylamine and nomifensine." Brain Res.564. 319-322 (1991)
• [文献書誌] Suzuki, T., Fujita, S., Narimatsu, S., Masubuchi, Y., Tachibana, M., Ohta, S.and Hirobe, M.: "Cytochrome P‐450 isozymes catalysing 4‐hydroxylation of parkinsonism related compound,1,2,3,4‐tetrahydroisoquinoline in rat liver microsomes." FASEB J.6. 771-776 (1992)
• [文献書誌] Yoshida, M., et al.: "The neuronal mechanism underlying parkinsonism and dyskinesia:differential roles of the putamen and caudate nucleus." Neurosci. Res.12. 31-40 (1991)
• [文献書誌] Yoshida, M., et al.: "Differential roles of the caudate nucleus and putamen in motor behavior of the cat as investigated by local injection of GABA antagonists." Neurosci. Res.10. 34-51 (1991)
• [文献書誌] Hayasaka, K., Ishida, T. and Horai, S.: "Heteroplasmy and polymorphisms in the major noncoding region of mitochindrial DNA in Japanese monkeys:association with tandemly repeated sequences." Mol. Biol. Evol.8. 399-415 (1991)
• [文献書誌] Goto, Y., Nonaka, I. and Horai, S.: "A new mtDNA mutation associated with mitochondrial myopathy,encephalopathy,lactic acidosis and stroke‐like episodes(MELAS)." Biochim. Biophys. Acta.1097. 238-240 (1991)
• [文献書誌] Miyabayashi, S., Hanamizu, H., Endo, H., Tada, K. and Horai, S.: "A new type of mitochondrial DNA deletion in patients with encephalomyopathy." J. Inher. Metab. Dis.14. 805-812 (1991)
• [文献書誌] Karasawa, N., Isomura, G., Yamada, K.and Nagatsu, I.: "Immunocytochemical localization of monoaminergic and non‐aminergic neurons in the house shrew(Suncus murinus)brain." Acta Histochem. Cytochem.24. 465-475 (1991)
• [文献書誌] Komori, K., Fujii, T., Karasawa, N., Yamada, K. and Nagatsu, I.: "Some heurons of the mouse cortex and caudoputamen contain aromatic L‐amino acid decarboxylase but not monoamines." Acta Histochem. Cytochem.24. 571-577 (1991)
• [文献書誌] Komori, K., Sakai, M., Karasawa, N., Yamada, K. and Nagatsu, I.: "Evidence for transient expression of tyrosine hydroxylase immunoreactivity in the mouse striatum and the effect of colchicine." Acta Histochem. Cytochem.24. 223-231 (1991)
• [文献書誌] Komori, K., Fujii, T. and Nagatsu, I.: "Do some tyrosine hydroxylase‐immunoreactive neurons in the human ventrolateral arcuate nucleus and globus pallidus produce only L‐DOPA?" Neurosci. Lett.133. 203-206 (1991)
• [文献書誌] Nagatsu, I., Yamada, K., Karasawa, N., Sakai, M., Takeuchi, T., Kaneda, N., Sasaoka, T., Katsuki, M., Fujita, K.and Nagatsu, T.: "Expression in brain sensory neurons of the transgene in transgenic mice carrying human tyroshine hydroxylase gene." Neurosci. Lett.127. 91-95 (1991)
• [文献書誌] Uehara, K., et al.: "Highly sensitive chemiluminescence method for determining myeloperoxidase in human polymorphonuclear leudocytes." Anal. Biochem.199. 191-196 (1991)
• [文献書誌] Ishida, M., et al.: "Changes in preference for receptor subtypes of congfigurational variants of a glutamate analog:conversion from the NMDA‐type to the non‐NMDA type." Brain Res.550. 152-156 (1991)
• [文献書誌] Shimamoto, K., et al.: "Synthesis of four diastereomeric L‐2‐(carboxycyclopropyl)glycines. Conformationally constrained L‐glutamate analogues." J. Org. Chem.56. 4167-4176 (1991)
• [文献書誌] Ishida, M., et al.: "Novel Kainate derivatives:potent depolarizing actions on spinal motoneurones and dosal root fibres in newborn rats." Brit. J. Pharmacol.104. 873-878 (1991)
• [文献書誌] Kudo, Y., et al.: "A significant increase in intracellular Ca^<2+> concentration induced by [2S,3R,4S]‐2‐[carboxycyclopropyl]glycine,a new potent NMDA agonist,in cultured rat hippocampal neurons." Brain Res.567. 342-345 (1991)
• [文献書誌] Ogawa, N., et al.: "Long‐lasting effect of ceruletide on dyskinesia and monoaminergic neuronal pathways in rats treated with iminodipropionitrile." Brain Res.556. 271-279 (1991)
• [文献書誌] Asanuma, M., et al.: "Distribution of beta‐2 adrenergic receptor messenger RNA in the rat brain by in situ hybridization hisstochemistry:effects of chronic reserpine treatment." Neurochem. Res.16. 1253-1256 (1991)
• [文献書誌] Haba, K., et al.: "Time course of changes in lipid peroxidation,pre‐and post‐synaptic cholinergic indices,NMDA receptor binding and neuronal death in gerbil hippocampus following transient ischemia." Brain Res.540. 116-122 (1991)
• [文献書誌] Kirino, T., Robinson, H.P.C., Miwa, A., Tamura, A. and Kawai, N.: "Disturbance of membrane function preceeding ischemic delayed neuronal death in the gerbil hippocampus." J. Cereb. Blood Flow Metab.
• [文献書誌] Kirino, T., Tsujita, Y. and Tamura, A.: "Induced tolerance to ischemia in gerbil hippocampal neurons." J. Cereb. Blood Flow Metab.11. 299-307 (1991)
• [文献書誌] Mizuno, Y.: "Key Topics in Brain Resarch,Parkinson's Disease.From Clinical Aspects to Molecular Basis,Nagatsu,T.,Narabayashi,H.and Yoshida,M.,eds." Springer‐Verlag, Wien/New York, 220 (1991)
• [文献書誌] Mizuno, Y., Ikebe, S., Hattori, N., Tanaka, M., Ohta, S., Kondo, T., Sato, T., Kagawa, Y. and Ozawa, T.: "Progress in Neuropathology Vol.7,Mitochondrial Encephalomyopathies.,Sato,T.and DiMauro,S.,eds." Raven Press,New York, 258 (1991)
• [文献書誌] Ohta, S., Tasaki, Y., Makino, Y., Tachikawa, O., Ohta, S. and Hirobe, M.: "Basic,Clinical and Therapeutic Aspects of Alzheimer's and Parkinson's Diseases,Vol.1,Nagatsu,T.,Fisher,A.and Yoshida,M.,eds." Plenum Press,New York, 808 (1991)
• [文献書誌] Makino, Y., Tasaki, Y., Kashiwasake, M., Tachikawa, O., Ohta, S. and Hirobe, M.: "Basic,Clinical and Therapeutic Aspects of Alzheimer's and Parkinson's Diseases,Vol.1,Nagatsu,T.,Fisher,A.and Yoshida,M.,eds." Plenum Press,New York, 808 (1991)
• [文献書誌] Ohta, S., Kohno, M., Makino, Y., Tachikawa, O., Tasaki, Y., Kikuchi, K. and Hirobe, M.: "Key Topics in Brain Resarch,Parkinson's Disease.From Clinical Aspects to Molecular Basis,Nagatsu,T.,Narabayashi,H.and Yoshida,M.,eds." Springer‐Verlag,Wien/New York, 220 (1991)
• [文献書誌] Yoshida, M., Ogawa, M. and Nagatsu, T.: "Key Topics in Brain Resarch,Parkinson's Disease.From Clinical Aspects to Molecular Basis,Nagatsu,T.,Narabayashi,H.and Yoshida,M.,eds." Springer‐Verlag,Wien/New York, 220 (1991)
• [文献書誌] Miyabayashi, S., Hanamizu, H., Endo, H., Tada, K. and Horai, S.: "Progress in Neuropathology Vol.7,Mitochondrial Encephalomyopathies.,Sato,T.and DiMauro,S.,eds." Raven Press,New York, 258 (1991)