1994 年度実績報告書

ヒトカテコールアミン生合成酵素遺伝子導入トランスジェニックマウスの組織化学的解析

研究課題

研究課題/領域番号	06454696
研究機関	藤田保健衛生大学
研究代表者	永津郁子藤田保健衛生大学, 医学部・解剖学, 教授 (80084573)
研究分担者	唐沢延幸藤田保健衛生大学医学部, 解剖学, 研究技術員 (70148287) 酒井正雄藤田保健衛生大学医学部, 解剖学, 講師 (90196048) 新井良八藤田保健衛生大学医学部, 解剖学, 助教授 (20159487) 山田啓喜藤田保健衛生大学衛生学部, 解剖学, 助教授 (60084604)
キーワード	カテコールアミン生合成酵素 / 遺伝子導入 / トランスジェニックマウス / 免疫組織化学 / ドーパミン / ノルアドレナリン / アドレナリン / セロトニン
研究概要	ヒトチロシン水酸化酵素遺伝子導入トランスジェニックマウスの組織化学的解析 1)ヒトチロシン水酸化酵素(hTH)遺伝子の5上流領域の5.0,2.5,0.2kbのDNA断片に、レポーター遺伝子として大腸菌クロラムフェニコールアセチル基転移酵素(CAT)遺伝子を連結した融合遺伝子を導入したトランスジェニック(Tg)マウス(TC50,TC25,TC02マウス)を作成して、hTH遺伝子のプロモーターの解析を行なった(Nagatsu et al.,J.Neural Transmission,96:85-104,1994)。また導入遺伝子(CAT)産物の存在を、Tgマウス(TC50マウス)の中脳腹側被蓋野の細胞と樹状突起、およびその神経突起の終末、扁桃体中心核に、免疫電顕で証明した(Arai et al.,Neurosci.Lett.,168:76-80,1994)。 2)L-DOPAに対する特異抗体を自家作成した(Karasawa et al.,Neurosci.Lett.,179:65-70,1994)ので、従来の研究で私達が作製してきたチロシン水酸化酵素(TH)、芳香族L-アミノ酸脱炭酸酵素(AADC)、ドーパミン-β-水酸化酵素(DBH)(1:5000)(Nagatsu et al.,Neurosci.Lett.,120:141-145,1990)、フェニールエタノールアミン-N-メチル基転移酵素(PNMT)、ドーパミン、セロトニンなどの各種の特異抗体も用いて、免疫組織化学によって酵素とその生成物との共存を明らかにした。このために二重染色法を用いた(Arai et al.,Brain Res.,667:295-299,1994)。ヒトドーパミンβ-水酸化酵素遺伝子導入トランスジェニックマウスの解析 3)4kbのヒトDBH遺伝子プロモーター領域に完全長のヒトDBHcDNAを連結した融合遺伝子を持つトランスジェニックマウス(DDBマウス)を作成した。その脳と副腎のノルアドレナリンとアドレナリン合成細胞に、ヒトDBHを発現させた。4kbのDBHプロモーターはノルアドレナリン細胞とアドレナリン細胞に変換することが示唆された。(Kobayashi et al.,J.Biol.Chem.,269:29725-29731,1994)。

研究成果

(21件)

すべてその他

すべて文献書誌 (21件)

[文献書誌] I.Nagatsu,N.Karasawa,K.Yamada,M.Sakai,T.Fujii,T.Takeuchi,R.Arai,K.Kobayashi,T.Nagatsu.: "Exprssion of human tyrosine hydroxylase-chioramphenico acetyltransferasc(CAT)fusion gene in the brains of transgenicmice as examined by CAT immunocytochemistry." J.Neural Transmission. 96. 85-104 (1994)
[文献書誌] R.Arai,N.Karasawa,S.Deura,K.Kobayashi,T.Nagatsu,I.Nagatsu.: "Protein products of the bacterial reporter gene are found within axon terminals in the brain of transgenic mice." Neurosci.Lett.168(1,2). 76-80 (1994)
[文献書誌] N.Karasawa,R.Arai,G.Isomura,K.Yamada,K.Sakai,M.Sakai,T.Nagatsu,I.Nagatsu.: "Phenotypic changes of AADC-only immunopositive premammillary neurons in the brain of iaboratory shrew (Suncus murinus)by systemic administration of monoamine precursors." Neurosci.Lett.179. 65-70 (1994)
[文献書誌] R.Arai,N.Karasawa,M.Geffard,T.Nagatsu,I.Nagatsu.: "Immunohistochemicalevidence that central serotonin neurons produce dopamine from exogenous L-DOPA in the rat,with reference to the involvement of aromatic L-amino aciddecarboxylase." Brain Res.667. 295-299 (1994)
[文献書誌] K.Kobayashi,S.Morita,T.Mizuguchi,H.Sawada,K.Yamada,I.Nagatsu,K.Fujita,T.Nagatsu.: "Functional and high-level expression of human dopamine β-hydroxylase in transgenicmice." J.Biolgical Chemistry. 269(47). 29725-29731 (1994)
[文献書誌] T.Fujii,M.Sakai,I.Nagatsu.: "Immunohistochemicaldemonstration of expression of tyrosine hydroxylase in ceredellar Purkinje cells of the human and mouse." Neurosci.Lett.165(1,2). 161-163 (1994)
[文献書誌] N.Karasawa,G.Isomura,K.Yamada,K.Sakai,I.Nagatsu.: "L-DOPA immunoreactiveneurons in the ventrolateralarea of arcuatenucleus of the house-shrew(Suncus murinus)." Biogenic Amines. 10(4). 287-293 (1994)
[文献書誌] N.Karasawa,R.Arai,G.Isomura,K.Yamada,M.Sakai,T.Nagatsu.I.Nagatsu.: "D-neurons(TH-negative,AADC-positive neurons)may belong to APUD system in the laboratory shrew(Suncus murinus)brain." Biogenic Amines. 10(4). 311-318 (1994)
[文献書誌] M.Tanaka,H.Okamura,Y.Tamada,I.Nagatsu,Y.Tanaka,Y.Ibata.: "Catecholaminergic input to spinally projecting serotonin neurons in the rostral ventromedial medullaoblongata of the rat." Brain Research Bulletin. 35(1). 23-30 (1994)
[文献書誌] M.Suzuki,Y.Ohmori,T.Watanabe,I.Nagatsu,: "Immunohistochemicalstudies on the intestinal nerve of Remak in the male chicken." J.Autonomic Nervous System. 49. 207-216 (1994)
[文献書誌] H.K.Inoue,A.Henschen,I.Nagatsu,L.Olson.: "Locus coeruleusterminals in intraocularly transplantedspinal cords as compared with catecholamine terminals in normal spinal cords:Thcir synaptic dcnsities and functional considerations." Med.Electron Microsci.27(2). 123-135 (1994)
[文献書誌] K.Inoue,C.R.Creveling,N.Karasawa,G.Isomura,I.Nagatsu.: "Measurementof DOPA and immunolocalization of L-DOPA-positive nerve fibers in rat dentalpulp." Brain Res.657. 307-309 (1994)
[文献書誌] K.Imamoto,K.Karasawa,G.Isomura,I.Nagatsu.: "Cajal-Retziusneurons idetified by GABA immunohistochemistry in layer I of the nat cerebralcortex." Neurosci.Res.20(1). 101-105 (1994)
[文献書誌] K.Iwase,A.Nagasaka,I.Nagatsu,K.Kiuchi,T.Nagatsu,H.Funahashi,T.Tsujimura,A.Inagaki,A.Nakai,T.Kishikawa,K.Miura.: "Tyrosine hydroxylase indicates cell diferentiation of catecholamine biosynthesis in neuroendocrine tumors." J.Endocrinol.Invest.17. 235-239 (1994)
[文献書誌] A.Monji,N.Morimoto,I.Okuyama,K.Umeno,I.Nagatsu,Y.Ibata,N.Tashiro.: "The number of noradrenergic and adrenergic neurons in the brain stem does not change with agein male Sprague-Dawley rats." Brain Res.641. 171-175 (1994)
[文献書誌] R.Arai,N.Karasawa,T.Nagatsu,I.Nagatsu.: "Exogenous L-5-hydroxytryptophan is decarboxylatec in neurons of the substantia nigra pars compacta and locus coeruleusof the rat." Brain Res.669. 145-149 (1995)
[文献書誌] R.Arai,D.M.Jacobowitz,I.Nagatsu.: "UP-regulation of calretininin the supraoptic nucleus of the rat after chronic salt loading." Brain Res.
[文献書誌] Y.Ichitani,H.Okamura,D.Nakahara.I.Nagatsu,Y.Ibata.: "Biochemicaland immunocytochemical changesinducedby intrastriata16-hydroxydopamine injection in the rat nigrostriatal dopamine neuron system:evidence for cell deathin the substantia nigra." Experimental Neurology.
[文献書誌] I.Nagatsu,K.Yamada,M.Sakai,N.Karasawa.: "Immunocytochemistry and in situ hybridization of catecholamine-synthesizing enzymes and the related neurotransmitters." In.“Methodsin Neurotransmitter and Neuropeptide Rescarch"(eds.Parvez,S.H.,Naoi,M.,Nagatsu,T.,Parvez,S.)Elsevier Science Publishers B.V.,Amsterdam., 151-183 (1993)
[文献書誌] I.Nagatsu,K.Yamaga,N.Karasawa,N.Kaneda,T.Sasaoka,K.Kobayashi,K.Fujita,T.Nagatsu.: "Non-catecholaminergicneuronal expression of hnman tyrosine hydroxylase in the brain of transgenic mice with special reference to aromatic L-amino acid decarboxylase." In“Tyrosine Hydroxylase"(eds.M.Naoi,S.H.Parvez)VSP Science Press,Utrecht., 37-57 (1993)
[文献書誌] K.Kitahama,I.Nagatsu,J.Pearson.: "Catecholaminesystems in mammalian midbrain and hindbrain:theme and variations." In“Phylogeny and Development of Catecholamine Systems in the CNS of Vertebrates"(eds.Smeets,W.J.A.J.,Reiner,A.)Cambridgc University Press,Cambridge., 183-205 (1994)

1994 年度 実績報告書

ヒトカテコールアミン生合成酵素遺伝子導入トランスジェニックマウスの組織化学的解析

研究代表者

永津 郁子 藤田保健衛生大学, 医学部・解剖学, 教授 (80084573)

研究成果

[文献書誌] R.Arai,N.Karasawa,S.Deura,K.Kobayashi,T.Nagatsu,I.Nagatsu.: "Protein products of the bacterial reporter gene are found within axon terminals in the brain of transgenic mice." Neurosci.Lett.168(1,2). 76-80 (1994)

[文献書誌] N.Karasawa,R.Arai,G.Isomura,K.Yamada,K.Sakai,M.Sakai,T.Nagatsu,I.Nagatsu.: "Phenotypic changes of AADC-only immunopositive premammillary neurons in the brain of iaboratory shrew (Suncus murinus)by systemic administration of monoamine precursors." Neurosci.Lett.179. 65-70 (1994)

[文献書誌] R.Arai,N.Karasawa,M.Geffard,T.Nagatsu,I.Nagatsu.: "Immunohistochemicalevidence that central serotonin neurons produce dopamine from exogenous L-DOPA in the rat,with reference to the involvement of aromatic L-amino aciddecarboxylase." Brain Res.667. 295-299 (1994)

[文献書誌] K.Kobayashi,S.Morita,T.Mizuguchi,H.Sawada,K.Yamada,I.Nagatsu,K.Fujita,T.Nagatsu.: "Functional and high-level expression of human dopamine β-hydroxylase in transgenicmice." J.Biolgical Chemistry. 269(47). 29725-29731 (1994)

[文献書誌] T.Fujii,M.Sakai,I.Nagatsu.: "Immunohistochemicaldemonstration of expression of tyrosine hydroxylase in ceredellar Purkinje cells of the human and mouse." Neurosci.Lett.165(1,2). 161-163 (1994)

[文献書誌] N.Karasawa,G.Isomura,K.Yamada,K.Sakai,I.Nagatsu.: "L-DOPA immunoreactiveneurons in the ventrolateralarea of arcuatenucleus of the house-shrew(Suncus murinus)." Biogenic Amines. 10(4). 287-293 (1994)

[文献書誌] N.Karasawa,R.Arai,G.Isomura,K.Yamada,M.Sakai,T.Nagatsu.I.Nagatsu.: "D-neurons(TH-negative,AADC-positive neurons)may belong to APUD system in the laboratory shrew(Suncus murinus)brain." Biogenic Amines. 10(4). 311-318 (1994)

[文献書誌] M.Tanaka,H.Okamura,Y.Tamada,I.Nagatsu,Y.Tanaka,Y.Ibata.: "Catecholaminergic input to spinally projecting serotonin neurons in the rostral ventromedial medullaoblongata of the rat." Brain Research Bulletin. 35(1). 23-30 (1994)

[文献書誌] M.Suzuki,Y.Ohmori,T.Watanabe,I.Nagatsu,: "Immunohistochemicalstudies on the intestinal nerve of Remak in the male chicken." J.Autonomic Nervous System. 49. 207-216 (1994)

[文献書誌] H.K.Inoue,A.Henschen,I.Nagatsu,L.Olson.: "Locus coeruleusterminals in intraocularly transplantedspinal cords as compared with catecholamine terminals in normal spinal cords:Thcir synaptic dcnsities and functional considerations." Med.Electron Microsci.27(2). 123-135 (1994)

[文献書誌] K.Inoue,C.R.Creveling,N.Karasawa,G.Isomura,I.Nagatsu.: "Measurementof DOPA and immunolocalization of L-DOPA-positive nerve fibers in rat dentalpulp." Brain Res.657. 307-309 (1994)

[文献書誌] K.Imamoto,K.Karasawa,G.Isomura,I.Nagatsu.: "Cajal-Retziusneurons idetified by GABA immunohistochemistry in layer I of the nat cerebralcortex." Neurosci.Res.20(1). 101-105 (1994)

[文献書誌] K.Iwase,A.Nagasaka,I.Nagatsu,K.Kiuchi,T.Nagatsu,H.Funahashi,T.Tsujimura,A.Inagaki,A.Nakai,T.Kishikawa,K.Miura.: "Tyrosine hydroxylase indicates cell diferentiation of catecholamine biosynthesis in neuroendocrine tumors." J.Endocrinol.Invest.17. 235-239 (1994)

[文献書誌] A.Monji,N.Morimoto,I.Okuyama,K.Umeno,I.Nagatsu,Y.Ibata,N.Tashiro.: "The number of noradrenergic and adrenergic neurons in the brain stem does not change with agein male Sprague-Dawley rats." Brain Res.641. 171-175 (1994)

[文献書誌] R.Arai,N.Karasawa,T.Nagatsu,I.Nagatsu.: "Exogenous L-5-hydroxytryptophan is decarboxylatec in neurons of the substantia nigra pars compacta and locus coeruleusof the rat." Brain Res.669. 145-149 (1995)

[文献書誌] R.Arai,D.M.Jacobowitz,I.Nagatsu.: "UP-regulation of calretininin the supraoptic nucleus of the rat after chronic salt loading." Brain Res.

[文献書誌] Y.Ichitani,H.Okamura,D.Nakahara.I.Nagatsu,Y.Ibata.: "Biochemicaland immunocytochemical changesinducedby intrastriata16-hydroxydopamine injection in the rat nigrostriatal dopamine neuron system:evidence for cell deathin the substantia nigra." Experimental Neurology.

[文献書誌] K.Kitahama,I.Nagatsu,J.Pearson.: "Catecholaminesystems in mammalian midbrain and hindbrain:theme and variations." In“Phylogeny and Development of Catecholamine Systems in the CNS of Vertebrates"(eds.Smeets,W.J.A.J.,Reiner,A.)Cambridgc University Press,Cambridge., 183-205 (1994)

1994 年度実績報告書

永津郁子藤田保健衛生大学, 医学部・解剖学, 教授 (80084573)