神経細胞死のメカニズムと防御に関する研究

• Project/Area Number: 03263105
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Research Institution: Juntendo University
• Principal Investigator: 水野 美邦 順天堂大学, 医学部・神経学講座, 教授 (30049043)
• Co-Investigator(Kenkyū-buntansha): 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)
• Project Period (FY): 1991 – 1992
• Project Status: Completed (Fiscal Year 1991)
• Budget Amount: ¥81,000,000 (Direct Cost: ¥81,000,000); Fiscal Year 1991: ¥81,000,000 (Direct Cost: ¥81,000,000)
• Keywords: 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

Research Abstract

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.

Research Products

• [Publications] 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)
• [Publications] Hattori, Y., Yoshino, H., Kondo, T., Mizuno, Y. and Horai, S.: "Is Parkinson's disease a“mitochondrial disease?"." J. Neurol. Sci.(1992)
• [Publications] 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)
• [Publications] 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)
• [Publications] Ito, H., et al.: "Mitochondrial leucine tRNA mutation in a mitochondrial encephalomyopathy." Lancet. 337. 234-235 (1991)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] Hayakawa, M., et al.: "Age‐associated accumulation of 8‐hydroxydeoxyguanosine in mitochondrial DNA of human diaphragm." Biochem. Biophys. Res. Commun.179. 1023-1029 (1991)
• [Publications] Ohno, K., et al.: "Direct DNA sequencing from colony:analysis of multiple deletions of mitochondrial genome." Biochim. Biophys. Acta. 1090. 9-16 (1991)
• [Publications] Nakamura, S., et al.: "The histochemical demonstration of monoamine oxidase‐containing neurons in the human hypothalamus." Neuroscience. 44. 457-463 (1991)
• [Publications] 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)
• [Publications] 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.
• [Publications] 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)
• [Publications] 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)
• [Publications] 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.
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] Uehara, K., et al.: "Highly sensitive chemiluminescence method for determining myeloperoxidase in human polymorphonuclear leudocytes." Anal. Biochem.199. 191-196 (1991)
• [Publications] 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)
• [Publications] Shimamoto, K., et al.: "Synthesis of four diastereomeric L‐2‐(carboxycyclopropyl)glycines. Conformationally constrained L‐glutamate analogues." J. Org. Chem.56. 4167-4176 (1991)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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.
• [Publications] Kirino, T., Tsujita, Y. and Tamura, A.: "Induced tolerance to ischemia in gerbil hippocampal neurons." J. Cereb. Blood Flow Metab.11. 299-307 (1991)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)
• [Publications] 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)