1989 年度実績報告書

カテコ-ルアミン関連酵素DNAプロ-ブによる遺伝性精神神経疾患に遺伝子解析

研究課題

研究課題/領域番号	01440047
研究機関	名古屋大学
研究代表者	永津俊治名古屋大学, 医学部, 教授 (40064802)
研究分担者	萩原昌子名古屋大学, 医学部, 教務員一瀬宏名古屋大学, 医学部, 助手 (90192492) 金田典雄名古屋大学, 医学部, 講師 (00144139) 直井信名古屋大学, 医学部, 助教授 (50022786)
キーワード	ヒト・チロシン水酸化酵素 / ヒト・ド-パミンβー水酸化酵素 / ヒト・フェニルエタノ-ルアミンNーメチル基転移酵素 / ヒト・ド-パ脱炭酸酵素 / cDNA / DNAプロ-ブ / カテコ-ルアミン生合成酵素 / トランスジェニックマウス
研究概要	本研究は、カテコ-ルアミン系神経伝達物質(ド-パミン、ノルアドレナリン、アドレナリン)のヒトの生合成酵素のDNAプロ-ブを用いて、遺伝性精神・神経疾患の発症の分子機構の解明を目指している。これまでに、クロ-ニングしたヒト・チロシン水酸化酵素(hTH)、ヒト・ド-パミンβー水酸化酵素、ヒト・フェニルエタノ-ルアミンNーメチル基転移酵素のDNAプロ-ブを用いて、トランスジェニックスマウスによるパ-キンソン病やそううつ病のモデル動物の確立を試みる。我々は、ヒト・ド-パ脱炭酸酵素(hDDC)のcDNAをクロ-ニングすることに成功し、4種のカテコ-ルアミン生合成酵素全てのDNAプロ-ブをそろえることができた。hDDCをヒト褐色細胞腫から均一し、精製酵素をリジルエンドペプチダ-ゼで加水分解して、生じるペプチドフラグメントのアミノ酸配列を気相式アミノ酸シ-クエンサ-により決定した。得られたアミノ酸配列の一つの相補的な38塩基長のDNAを合成し、プロ-ブとし、6x10^5のヒト褐色細胞腫cDNAライブラリ-をスクリ-ニングすることにより、hDDCの全アミノ酸配列を規定している領域を含む完全長cDNAクロ-ンを単離した。単離されたcDNAは、全長1932塩基で、480アミノ酸からなる分子量53.891のタンパク質を規定していた。予想される補酵素ピリドキサルりん酸の結合部位周辺のアミノ酸配列(ーAsnーPheーAsnーProーHisーLysーTrpー)は、ヒト・ブタ・ハエで完全に一致していた。この部位のアミノ酸配列が、基質特異性の決定あるいは本酵素の活性発現に重要な役割を担っていることを意味する。また、hTHを導入したトランスジェニックマウスの作製にも成功しており,現在hTHの発現量の発現部位等について検討中である。家族性パ-キンソン病におけるhTHをプロ-ブとして用いた制限酵素断片長多型性の研究では、米国の患者において制限酵素断片の変化を見つけたので、家系分析を行なうための試料を採集中である。

研究成果

(36件)

すべてその他

すべて文献書誌 (36件)

[文献書誌] Tsuruta,M.: "Differences in the response if circulating noradrenaline and adrenaline in exerciseinduced asthma." Biogenic Amines.6. 75-80 (1989)
[文献書誌] Kobayashi,K.: "Human dopamine-hydroxylase gene:two mRNA types having different 3′-terminal regions are produced through alternative polyadenylation." Nucleic Acids Research.17. 1089-1102 (1989)
[文献書誌] Naoi,M.: "Oxidation of N-methyl-1,2,3,4-tetrahydroisoquinoline into the N-methyl-isoquinolinium ion by monoamine oxidase." J.Neurochem.52. 653-655 (1989)
[文献書誌] Naoi,M.: "Uptake of heterocyclic amines,Trp-P-1 and Trp-P-2,into clonal rat pheochromocytoma PC12h cells by dopamine uptake system." Neurosci.Lett.99. 317-322 (1989)
[文献書誌] Nagatsu,I.: "Quantitative analysis of reduction of aromatic-L-amino acid decarboxylase and tyrosine hydroxylase-like immunoreactivities in the nigrostriatal dopaminergic neurons of MPTP-treated mice." Biogenic Amines.6. 263-277 (1989)
[文献書誌] Nagatsu,T.: "Parkinson′s disease:Degenerative Brain Disease,A Molecular Biology Approach." Asian Med.J.32. 304-312 (1989)
[文献書誌] Gotoh,H.: "Activities of dipeptidyl peptidase II and dipeptidyl peptidase IV in synovial fluid from patients with rheumatoid arthritis and osteoarthritis." Clin.Chem.35. 1016-1018 (1989)
[文献書誌] Kozawa,T.: "Relationship between severity of asthma and response of noradrenaline and adrenaline to exercise." Biogenic Amines.9. 301-308 (1989)
[文献書誌] Naoi,M.: "A N-methyltransferase in human brain catalyses N-methylation of 1,2,3,4-tetrahydroisoquinoline into N-methyl-1,2,3,4-tetrahydroisoquinoline,a precursor of a dopaminergic neurotoxin,N-methylisoquinolinium ion." Biochem.Biophys.Res.Commun.161. 1213-1219 (1989)
[文献書誌] Niwa,T.: "Detection of tetrahydroisoquinoline in parkinsonian brain as an endogenous amine by use of gas chromatography-mass spectrometry." J.Chromatogr.491. 397-403 (1989)
[文献書誌] Niwa,T.: "Presence of tetrahydroisoquinoline,a parkinsonism-related compound,in foods." J.Chromatogr.493. 347-352 (1989)
[文献書誌] Hagihara,M.: "Cyclosporin A,an immune suppressor,enhanced neurotoxicity of N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to mice." Neurochem.Int.15. 249-254 (1989)
[文献書誌] Nakahara,D.: "Increased dopamine and serotonin metabolism in rat nucleus accumbens produced by intracranial self-stimulation of medial forebrain bundle as measured by in vivo microdialysis." Brain Res.495. 178-181 (1989)
[文献書誌] Uchida,K.: "Synthesis of L-3,4-dihydroxyphenylalanine by tyrosine hydroxylase cDNA-transfected C6 cells:application for intracerebral grafting." J.Neurochem.53. 728-732 (1989)
[文献書誌] Ogawa,M.: "The effect of 1,2,3,4-tetrahydroisoquinoline (TIQ)on mesencephalic dopaminergic neurons in C57BL/6J mice:immunohistochemical studiestyrosine hydroxylase." Biogenic Amines.6. 427-436 (1989)
[文献書誌] Magatsu,T.: "The human tyrosine hydroxylase gene." Molecular and Cellular Neuribiology.6. 313-321 (1989)
[文献書誌] Mogi.M.: "2-Microglobulin decrease in cerebrospinal fluid form parkinsonian patients." Neurosci.Lett.104. 241-246 (1989)
[文献書誌] Nagatsu,T.: "Parkinson′s diseaseーーa representative aging associated neurodegenerative disease." LEADERSHIP MEDICA.8. 4-10 (1989)
[文献書誌] Hagihara,M.: "Activities of dipeptidyl peptidases in BXSB mice and MRL/lpr mice with lupus erythematosus-like syndrome." Biochem.Med.Metab.Biol.42. 224-231 (1989)
[文献書誌] Kojima,T.: "3-amino-1-methyl-5H-pyrido[4,3ーb]indole (Trp-P-2)and other heterocyclic amines as inhibitors of mitochondrial monoamine oxidases separated from human brain synaptosomes." Neurochem.Int.16. 51-57 (1990)
[文献書誌] Ishii,A.: "Ferrous ion activates the less active form of human adrenal tyrosine hydroxylase." Neurochem.Int.16. 59-64 (1990)
[文献書誌] Ohya,Y.: "Uptake of N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)and the N-methyl-4-phenylpyridinium ion (MPP^+)into fetal mouse brain through the placenta." Neurosci.Lett.105. 221-226 (1989)
[文献書誌] Nakahara,D.: "A removable brain microdialysis probe unit for in vivo monitoring of neurochemical activity." Biogenic Amines.6. 559-564 (1989)
[文献書誌] Naoi,M.: "N-Methylisoquinolinium ion as an inhibitor of tyrosine hydroxylase,aromatic L-amino acid decarboxylase and monoamine oxidase." Neurochem.Int.15. 315-320 (1989)
[文献書誌] Mogi,M.: "Selective removal of beta_2-microglobulin from human plasma by highperformance immunoaffinity chromatography." J.Chromatogr.496. 194-200 (1989)
[文献書誌] Ichinose H.: "Isolation and characterization of a cDNA clone encoding human aromatic L-amino acid decarboxylase." Biochem.Biophys.Res.Commun.164. 1024-1030 (1989)
[文献書誌] Sasaoka,T.: "Structures of human phenylethanolamine N-methyltransferase gene:existence of two types of mRNA with different transcription initiation sites." Neurochem.Int.15. 555-565 (1989)
[文献書誌] Nagatsu,T.: "Abnormalcatecholamine metabolism in spontaneously hypertensiverats volammetry in vivo.In:Progress in hypertension,vol.1," H.Saito(Ed.). VNU Science Press,Utrecht,1-8 (1989)
[文献書誌] Nagatsu,T.: "Regulation of catecholamine synthesis by tetrahydrobiopterin as cofactor of tyrosine hydroxylase.In:Progress in Catecholamine Research," A.Dahlstrom,R.h.Belmaker and M.Sandler(Eds.) Alanb R. Liss,Inc.,New York,41-46 (1989)
[文献書誌] Nagatsu,T.: "Inhibitors and stimulatorsnof monoamine-synthesizing enzymes from microbes.In:Horizons on Antibiotic Research," B.D.Davis et al.(Eds.).Japan Antibiotics Research Association,Tokyo,121-137 (1989)
[文献書誌] Nagatsu,T.: "Physiological and clinical chemistry of biopterin.In:Medical Research Reviews," G.deStevens(Ed.).John Wiley & Suns,New York,25-44 (1989)
[文献書誌] Nagatsu,T.: "Molecular genetic approaches to the study of human catecholaminesynthesizing enzymes.In:Bioinformatics," O.Hatase,J.H.Wang(Eds.).Elsevier Science Publishers B.V.,Amsterdam,281-288 (1989)
[文献書誌] Kojima,K.: "Microbore HPLC for biological samples:catecholamines,peptides and proteins.In:PROGRESS IN HPLC,vol.4," M.Yoshioka,S.Parvez,H.Parvez.(Eds.).VNU Science Press,Utrecht,211-227 (1989)
[文献書誌] Nagatsu,T.: "Changes of brain monoamines under stress measured in vivo by voltammetry.In:Stree:Neurochemical and Humoral Mechanisms," G.R.Van Loon,R.Kvetnansky,R.McCarry and J.Axelrod(Eds.).Gordon and Breach Science,New York,43-51 (1989)
[文献書誌] Nagatsu,T.: "Roles of calcium ion in the regulation of tyrosine hydroxylase.In:Calcium Protein signaling," H.Hidaka,E.Carafoli,A.R.Means and T.Tanaka(Eds.)Plenum Publishing Corporation,New York,425-433 (1989)
[文献書誌] Nakajima,T.: "Neuron-specific enolase and aromatic L-amino acid decarboxylase;neuroendocrine makers in immunohistochemistry.In:Endocrine Pathology update," J.Lechago and T.Kameya(Eds.).Neuroendocrine Markers,Field and Wood,Inc.,Philadelphia,277-292 (1989)

1989 年度 実績報告書

カテコ-ルアミン関連酵素DNAプロ-ブによる遺伝性精神神経疾患に遺伝子解析

研究代表者

永津 俊治 名古屋大学, 医学部, 教授 (40064802)

研究成果

[文献書誌] Tsuruta,M.: "Differences in the response if circulating noradrenaline and adrenaline in exerciseinduced asthma." Biogenic Amines.6. 75-80 (1989)

[文献書誌] Kobayashi,K.: "Human dopamine-hydroxylase gene:two mRNA types having different 3′-terminal regions are produced through alternative polyadenylation." Nucleic Acids Research.17. 1089-1102 (1989)

[文献書誌] Naoi,M.: "Oxidation of N-methyl-1,2,3,4-tetrahydroisoquinoline into the N-methyl-isoquinolinium ion by monoamine oxidase." J.Neurochem.52. 653-655 (1989)

[文献書誌] Naoi,M.: "Uptake of heterocyclic amines,Trp-P-1 and Trp-P-2,into clonal rat pheochromocytoma PC12h cells by dopamine uptake system." Neurosci.Lett.99. 317-322 (1989)

[文献書誌] Nagatsu,I.: "Quantitative analysis of reduction of aromatic-L-amino acid decarboxylase and tyrosine hydroxylase-like immunoreactivities in the nigrostriatal dopaminergic neurons of MPTP-treated mice." Biogenic Amines.6. 263-277 (1989)

[文献書誌] Nagatsu,T.: "Parkinson′s disease:Degenerative Brain Disease,A Molecular Biology Approach." Asian Med.J.32. 304-312 (1989)

[文献書誌] Gotoh,H.: "Activities of dipeptidyl peptidase II and dipeptidyl peptidase IV in synovial fluid from patients with rheumatoid arthritis and osteoarthritis." Clin.Chem.35. 1016-1018 (1989)

[文献書誌] Kozawa,T.: "Relationship between severity of asthma and response of noradrenaline and adrenaline to exercise." Biogenic Amines.9. 301-308 (1989)

[文献書誌] Naoi,M.: "A N-methyltransferase in human brain catalyses N-methylation of 1,2,3,4-tetrahydroisoquinoline into N-methyl-1,2,3,4-tetrahydroisoquinoline,a precursor of a dopaminergic neurotoxin,N-methylisoquinolinium ion." Biochem.Biophys.Res.Commun.161. 1213-1219 (1989)

[文献書誌] Niwa,T.: "Detection of tetrahydroisoquinoline in parkinsonian brain as an endogenous amine by use of gas chromatography-mass spectrometry." J.Chromatogr.491. 397-403 (1989)

[文献書誌] Niwa,T.: "Presence of tetrahydroisoquinoline,a parkinsonism-related compound,in foods." J.Chromatogr.493. 347-352 (1989)

[文献書誌] Hagihara,M.: "Cyclosporin A,an immune suppressor,enhanced neurotoxicity of N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to mice." Neurochem.Int.15. 249-254 (1989)

[文献書誌] Nakahara,D.: "Increased dopamine and serotonin metabolism in rat nucleus accumbens produced by intracranial self-stimulation of medial forebrain bundle as measured by in vivo microdialysis." Brain Res.495. 178-181 (1989)

[文献書誌] Uchida,K.: "Synthesis of L-3,4-dihydroxyphenylalanine by tyrosine hydroxylase cDNA-transfected C6 cells:application for intracerebral grafting." J.Neurochem.53. 728-732 (1989)

[文献書誌] Ogawa,M.: "The effect of 1,2,3,4-tetrahydroisoquinoline (TIQ)on mesencephalic dopaminergic neurons in C57BL/6J mice:immunohistochemical studiestyrosine hydroxylase." Biogenic Amines.6. 427-436 (1989)

[文献書誌] Magatsu,T.: "The human tyrosine hydroxylase gene." Molecular and Cellular Neuribiology.6. 313-321 (1989)

[文献書誌] Mogi.M.: "2-Microglobulin decrease in cerebrospinal fluid form parkinsonian patients." Neurosci.Lett.104. 241-246 (1989)

[文献書誌] Nagatsu,T.: "Parkinson′s diseaseーーa representative aging associated neurodegenerative disease." LEADERSHIP MEDICA.8. 4-10 (1989)

[文献書誌] Hagihara,M.: "Activities of dipeptidyl peptidases in BXSB mice and MRL/lpr mice with lupus erythematosus-like syndrome." Biochem.Med.Metab.Biol.42. 224-231 (1989)

[文献書誌] Kojima,T.: "3-amino-1-methyl-5H-pyrido[4,3ーb]indole (Trp-P-2)and other heterocyclic amines as inhibitors of mitochondrial monoamine oxidases separated from human brain synaptosomes." Neurochem.Int.16. 51-57 (1990)

[文献書誌] Ishii,A.: "Ferrous ion activates the less active form of human adrenal tyrosine hydroxylase." Neurochem.Int.16. 59-64 (1990)

[文献書誌] Ohya,Y.: "Uptake of N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)and the N-methyl-4-phenylpyridinium ion (MPP^+)into fetal mouse brain through the placenta." Neurosci.Lett.105. 221-226 (1989)

[文献書誌] Nakahara,D.: "A removable brain microdialysis probe unit for in vivo monitoring of neurochemical activity." Biogenic Amines.6. 559-564 (1989)

[文献書誌] Naoi,M.: "N-Methylisoquinolinium ion as an inhibitor of tyrosine hydroxylase,aromatic L-amino acid decarboxylase and monoamine oxidase." Neurochem.Int.15. 315-320 (1989)

[文献書誌] Mogi,M.: "Selective removal of beta_2-microglobulin from human plasma by highperformance immunoaffinity chromatography." J.Chromatogr.496. 194-200 (1989)

[文献書誌] Ichinose H.: "Isolation and characterization of a cDNA clone encoding human aromatic L-amino acid decarboxylase." Biochem.Biophys.Res.Commun.164. 1024-1030 (1989)

[文献書誌] Sasaoka,T.: "Structures of human phenylethanolamine N-methyltransferase gene:existence of two types of mRNA with different transcription initiation sites." Neurochem.Int.15. 555-565 (1989)

[文献書誌] Nagatsu,T.: "Abnormalcatecholamine metabolism in spontaneously hypertensiverats volammetry in vivo.In:Progress in hypertension,vol.1," H.Saito(Ed.). VNU Science Press,Utrecht,1-8 (1989)

[文献書誌] Nagatsu,T.: "Regulation of catecholamine synthesis by tetrahydrobiopterin as cofactor of tyrosine hydroxylase.In:Progress in Catecholamine Research," A.Dahlstrom,R.h.Belmaker and M.Sandler(Eds.) Alanb R. Liss,Inc.,New York,41-46 (1989)

[文献書誌] Nagatsu,T.: "Inhibitors and stimulatorsnof monoamine-synthesizing enzymes from microbes.In:Horizons on Antibiotic Research," B.D.Davis et al.(Eds.).Japan Antibiotics Research Association,Tokyo,121-137 (1989)

[文献書誌] Nagatsu,T.: "Physiological and clinical chemistry of biopterin.In:Medical Research Reviews," G.deStevens(Ed.).John Wiley & Suns,New York,25-44 (1989)

[文献書誌] Nagatsu,T.: "Molecular genetic approaches to the study of human catecholaminesynthesizing enzymes.In:Bioinformatics," O.Hatase,J.H.Wang(Eds.).Elsevier Science Publishers B.V.,Amsterdam,281-288 (1989)

[文献書誌] Kojima,K.: "Microbore HPLC for biological samples:catecholamines,peptides and proteins.In:PROGRESS IN HPLC,vol.4," M.Yoshioka,S.Parvez,H.Parvez.(Eds.).VNU Science Press,Utrecht,211-227 (1989)

[文献書誌] Nagatsu,T.: "Changes of brain monoamines under stress measured in vivo by voltammetry.In:Stree:Neurochemical and Humoral Mechanisms," G.R.Van Loon,R.Kvetnansky,R.McCarry and J.Axelrod(Eds.).Gordon and Breach Science,New York,43-51 (1989)

[文献書誌] Nagatsu,T.: "Roles of calcium ion in the regulation of tyrosine hydroxylase.In:Calcium Protein signaling," H.Hidaka,E.Carafoli,A.R.Means and T.Tanaka(Eds.)Plenum Publishing Corporation,New York,425-433 (1989)

[文献書誌] Nakajima,T.: "Neuron-specific enolase and aromatic L-amino acid decarboxylase;neuroendocrine makers in immunohistochemistry.In:Endocrine Pathology update," J.Lechago and T.Kameya(Eds.).Neuroendocrine Markers,Field and Wood,Inc.,Philadelphia,277-292 (1989)

1989 年度実績報告書

永津俊治名古屋大学, 医学部, 教授 (40064802)