2000 Fiscal Year Annual Research Report

延髄孤束核での一酸化窒素合成酵素の単一分子の動きの観察

Research Project

Project/Area Number	12670071
Research Institution	Wakayama Medical University
Principal Investigator	前田正信和歌山県立医科大学, 医学部, 教授 (80181593)
Co-Investigator(Kenkyū-buntansha)	真壁恭子和歌山県立医科大学, 医学部, 助手 (10203952) 坪田裕司和歌山県立医科大学, 医学部, 助手 (50197761) 湯川和典和歌山県立医科大学, 医学部, 講師 (20301434) 工藤秀明産業医科大学, 医学部, 講師 (40289575) 土肥良秋産業医科大学, 医学部, 助教授 (30258602)
Keywords	孤束核 / 一酸化窒素 / 一酸化窒素合成酵素 / green fluorescent protein(GFP) / 蛍光 / 脳 / 循環 / 遺伝子
Research Abstract	延髄孤束核(nucleus tractus solitarius,NTS)で一酸化窒素合成酵素(NO synthase,NOS)がどれくらいの頻度で合成されどれぐらいの時間で消滅していくかを調べるため、NTS内へ神経型NOS(neuronal NOS,nNOS)の遺伝子と、nNOSの遺伝子の下流にgreen fluorescent protein(GFP,染色しなくともそれ自身が蛍光で光る蛋白質)の遺伝子を組み込んだベクターを導入することを目指し、以下の実験を行った。尚、研究代表者は平成12年9月に和歌山県立医科大学へ転任したため、最初は、中枢性循環調節の研究が行える実験室にするための整備から始めた。 1.先ず、実験室を実体顕微鏡下にラットの定位脳手術の行えるように整備し、次に、血圧、心拍数等の循環機能のモニターができるように整えた。 2.nNOSのクローニングはSnyderら(Nature,1991)によって既に行われている。そこで、Snyderらに連絡をとり、クローニングされたnNOSの遺伝子を送ってもらい、それを使うこととした。 3.nNOSの遺伝子を組み込んでいないGFP Fusion TOPO Cloning Vectorsを作り、GFPを発現させ、それが蛍光を発することを確かめた。 4.次に、nNOSの遺伝子をGFP Fusion TOPO Cloning Vectors内へ組み込む実験を始めている。

Research Products

(20 results)

All Other

All Publications (20 results)

[Publications] Nakai,M.: "Cerebral cortical muscarinic cholinergic and N-methyl-1)-aspartate receptors mediate increase in cortieal blood flow elicited by chemical stimulation of periaqueductal gray matter."Neuroscience. 98(3). 449-457 (2000)
[Publications] Washio,M.: "The isolation of Methicillin-resistant Staphylococcus aureus (MRSA) is a predictor for poor prognosis of the elderly patients with bacterial infection."International Medical Journal,. 7(2). 117-120 (2000)
[Publications] Maeda,M.: "International comparison of periodic health examination and medical privacy of workers."Journal of UOEH. 22(Suppl). 165-168 (2000)
[Publications] Inoue,M: "Effect cf different work styles on blood pressure."Journal of UOEH. 22(Suppl). 197-200 (2000)
[Publications] Yukawa,K.: "Impaired water-maze performance in mice lacking transcription factor CCAAT/enhancer-binding protein beta."Neuroscience Research Communications. 26. 59-67 (2000)
[Publications] Shi,W.: "The class IV semaphorin CD100 plays a nonredundunt role in the immune system : Defective B and T cell activation in CD100/sema4D-deficient mice."Immunity. 13. 633-642 (2000)
[Publications] Kudo,H.: "Dietary zinc deficiency decreases glutathione S-transferase expression in the rat olfactory epithelium"Journal of Nutrition. 130(1). 38-44 (2000)
[Publications] Tamura,M.: "Expression of profilin, an actin-binding protein, in rat experimental glomerulonephritis and its upregulation by basic fibroblast growth factor in cultured rat mesangial cells"J Am Soci Nephrology. 11(3). 423-433 (2000)
[Publications] Doi,Y.: "Expression of a-calcitonin gene-related peptide in the enteric nervous system of rat small intestine"Neuroscience Letters. 285(1). 33-36 (2000)
[Publications] Uchida,S.: "Transient expression of activin bA mRNA on osteoprogenitor cells in rat bone regeneration after drill-hole injury"Bone. 27(1). 81-90 (2000)
[Publications] Fukushige,H.: "Synthesis and receptor sites of endothelin-1 in the rat liver vasculature"Anatomical Record. 259(4). 437-445 (2000)
[Publications] Yamamoto,O.: "Sweat gland toxicity induced by bis (tributyltin) oxide : An ultrastructural and X-ray microanalysis study"Archives of Toxicology. 74. 627-631 (2000)
[Publications] Nishino,T.: "Immunocytochemistry of glutathione S-transferase in taste bud cells of rat circumvallate and foliate papillae."Chemical Senses. 26. 179-188 (2001)
[Publications] Doi,Y.: "Synthesis of calcitonin gene-realted peptide (CGRP) by rat arterial endothelial cells"Histology and Histopathology. (in press). (2001)
[Publications] Kiyonaga,H.: "Expressions of endothelin-1, fibronectin and interleukin-1a of human umbilical vein endothelial cells under prolinged culture"Medical Electron Microscopy. (in press). (2001)
[Publications] 前田正信: "アンチセンス法を用いて延髄孤束核のnNOSの遺伝子を制御した時の循環変化"自律神経. 37(3). 395-399 (2000)
[Publications] 井上正岩: "液体塩素による凍傷事例からみた労働衛生スタッフの役割"産業衛生学雑誌. 42(6). 226-227 (2000)
[Publications] 坪田裕司: "筋疾患マウスB6CBA/J-croの骨格筋小胞体分画における蛋白欠損とその生化学的性質に関する研究."和歌山医学. 51. 181-192 (2000)
[Publications] 真壁恭子: "免疫不全マウスにおける胸腺細胞分化異常の解析と分化の異常"和歌山医学. 52(2)(in press). (2001)
[Publications] Fujimoto,S.: "Roles of endothelin-1 in regulation of rat hepatic sinusoidal blood flow In : European Conference on Microcirculation, Fagrell, B.(ed.)"Monduzzi Editore, Bologna. 33-37 (2000)

2000 Fiscal Year Annual Research Report

延髄孤束核での一酸化窒素合成酵素の単一分子の動きの観察

Principal Investigator

前田 正信 和歌山県立医科大学, 医学部, 教授 (80181593)

Research Products

[Publications] Nakai,M.: "Cerebral cortical muscarinic cholinergic and N-methyl-1)-aspartate receptors mediate increase in cortieal blood flow elicited by chemical stimulation of periaqueductal gray matter."Neuroscience. 98(3). 449-457 (2000)

[Publications] Washio,M.: "The isolation of Methicillin-resistant Staphylococcus aureus (MRSA) is a predictor for poor prognosis of the elderly patients with bacterial infection."International Medical Journal,. 7(2). 117-120 (2000)

[Publications] Maeda,M.: "International comparison of periodic health examination and medical privacy of workers."Journal of UOEH. 22(Suppl). 165-168 (2000)

[Publications] Inoue,M: "Effect cf different work styles on blood pressure."Journal of UOEH. 22(Suppl). 197-200 (2000)

[Publications] Yukawa,K.: "Impaired water-maze performance in mice lacking transcription factor CCAAT/enhancer-binding protein beta."Neuroscience Research Communications. 26. 59-67 (2000)

[Publications] Shi,W.: "The class IV semaphorin CD100 plays a nonredundunt role in the immune system : Defective B and T cell activation in CD100/sema4D-deficient mice."Immunity. 13. 633-642 (2000)

[Publications] Kudo,H.: "Dietary zinc deficiency decreases glutathione S-transferase expression in the rat olfactory epithelium"Journal of Nutrition. 130(1). 38-44 (2000)

[Publications] Tamura,M.: "Expression of profilin, an actin-binding protein, in rat experimental glomerulonephritis and its upregulation by basic fibroblast growth factor in cultured rat mesangial cells"J Am Soci Nephrology. 11(3). 423-433 (2000)

[Publications] Doi,Y.: "Expression of a-calcitonin gene-related peptide in the enteric nervous system of rat small intestine"Neuroscience Letters. 285(1). 33-36 (2000)

[Publications] Uchida,S.: "Transient expression of activin bA mRNA on osteoprogenitor cells in rat bone regeneration after drill-hole injury"Bone. 27(1). 81-90 (2000)

[Publications] Fukushige,H.: "Synthesis and receptor sites of endothelin-1 in the rat liver vasculature"Anatomical Record. 259(4). 437-445 (2000)

[Publications] Yamamoto,O.: "Sweat gland toxicity induced by bis (tributyltin) oxide : An ultrastructural and X-ray microanalysis study"Archives of Toxicology. 74. 627-631 (2000)

[Publications] Nishino,T.: "Immunocytochemistry of glutathione S-transferase in taste bud cells of rat circumvallate and foliate papillae."Chemical Senses. 26. 179-188 (2001)

[Publications] Doi,Y.: "Synthesis of calcitonin gene-realted peptide (CGRP) by rat arterial endothelial cells"Histology and Histopathology. (in press). (2001)

[Publications] Kiyonaga,H.: "Expressions of endothelin-1, fibronectin and interleukin-1a of human umbilical vein endothelial cells under prolinged culture"Medical Electron Microscopy. (in press). (2001)

[Publications] 前田正信: "アンチセンス法を用いて延髄孤束核のnNOSの遺伝子を制御した時の循環変化"自律神経. 37(3). 395-399 (2000)

[Publications] 井上正岩: "液体塩素による凍傷事例からみた労働衛生スタッフの役割"産業衛生学雑誌. 42(6). 226-227 (2000)

[Publications] 坪田裕司: "筋疾患マウスB6CBA/J-croの骨格筋小胞体分画における蛋白欠損とその生化学的性質に関する研究."和歌山医学. 51. 181-192 (2000)

[Publications] 真壁恭子: "免疫不全マウスにおける胸腺細胞分化異常の解析と分化の異常"和歌山医学. 52(2)(in press). (2001)

[Publications] Fujimoto,S.: "Roles of endothelin-1 in regulation of rat hepatic sinusoidal blood flow In : European Conference on Microcirculation, Fagrell, B.(ed.)"Monduzzi Editore, Bologna. 33-37 (2000)

前田正信和歌山県立医科大学, 医学部, 教授 (80181593)