2000 Fiscal Year Annual Research Report

単一細胞機能評価の新手法

Research Project

Project/Area Number	11227202
Research Institution	Tokyo University of Agriculture and Technology
Principal Investigator	松岡英明東京農工大学, 工学部, 教授 (10143653)
Co-Investigator(Kenkyū-buntansha)	橋本壽正東京工業大学, 大学院・工学研究科, 教授 (50101001) 夏目雅裕東京農工大学, 農学部, 助教授 (10201683) 竹山春子東京農工大学, 工学部, 助教授 (60262234) 安藤譲二東京大学, 大学院・医学系研究科, 教授 (20159528) 本多裕之名古屋大学, 大学院・工学研究科, 助教授 (70209328)
Keywords	単一細胞機能 / 電気的シグナル / イネキチナーゼ / 流水せん断応力 / 電気化学的シグナル / 熱シグナル / 遺伝子発現制御 / Ca^<2+>
Research Abstract	本年度は各研究課題毎に次のような成果を挙げた。 (1)イネ培養細胞にCa^<2+>を機械的に注入することでキチナーゼ遺伝子が発現した。次に電気的シグナルでCa^<2+>の導入を試み、1mV×30secでCa^<2+>を流入させることができた。それによるキチナーゼ発現は、まだ観擦されていない。キチナーゼは複数のアイソザイムがあるが、その発現パターンを活性染色法で解析し、さらに各々のアイソザイムのアミノ酸配列解析さらにその遺伝子解析を進めた。 (2)血管内皮細胞に流水せん断応力を作用させるとCa^<2+>チャンネル(P2X4)が開き、Ca^<2+>が細胞内に流入するが電気的シグナル(10Vp-p,1.0MHz正弦波)によってもCa^<2+>流入が観測された。一方、P2X4の遺伝子の発現は流水せん断応力に対してダウンレギュレーションされることも明らかにされた。 (3)酵母細胞に電気化学的シグナルを印加(1.0Vvs.SCE)した。この条件で生存率は1%程度になった。この時、発現する遺伝子を遺伝子チップで解析した結果、ヒートショックタンパク(HSP)70ファミリーに属する遺伝子も含め30以上の遺伝子の発現誘導ないし促進が認められた。 (4)熱シグナルに対するガン細胞の治癒条件の解析、植物細胞中の熱拡散率解析などを実施し、定量的な結果を得た。 (5)放射菌気菌糸分化因子の作用機序に関して、Ca^<2+>を介した電気的分化制御の可能性を検討した。以上を総括すると、当初の計画していた研究目的は概ね達成された。

Research Products

(18 results)

All Other

All Publications (18 results)

[Publications] M.Saito: "pH microelectrode-micropipette system for the measurement of enzyme reactions in a picoliter volume of a living plant cell"Anal.Chim.Acta. 404(2). 223-229 (2000)
[Publications] M.Saito: "Elicitor action via cell membrane of a cultured rice cell demonstrated by the single-cell transient assay"J.Biotechnol.. 76(2/3). 227-232 (2000)
[Publications] M.Saito: "Modulation of Ca^<2+> influx into a single rice cell with an electric signal"Electrochemistry. 68(5). 333-336 (2000)
[Publications] M.Saito: "Use of acetylene for the fabrication of a glass capillary carbon microelec-trode"Electrochemistry. 68(11). 924-926 (2000)
[Publications] H.Matsuoka: "Microbioelectronics for single-cell experiment"Electrochemistry. 68(5). 314-320 (2000)
[Publications] 斉藤美佳子: "ボーダーレス電気化学者をめざす"電気化学. 69(2). 128-129 (2001)
[Publications] M.Natsume: "Effect of blue light on endogenous brassinosteroid activity and leaf unrolling in rice seedlings."ITE Letters on Batteroes, New Technologuies & Medicine. 1. 781-785 (2000)
[Publications] M.Natsume: "Teasterone-3-O-b-D-glucopyranoside, a new conjugated brassinosteroid metabolite from lily cell suspension cultures and its identification in lily anthors."Biosci.Biotech.Biochem.. 64. 702-709 (2000)
[Publications] M.Natsume: "Reversible Conversion between Teasterone and Its Ester Conjugates in Lily Cell Culture."J.Pesticide Sci.. 25. 117-122 (2000)
[Publications] M.Natsume: "Effects of De-N-methylation of Pamamycins on Aerial Mycelium-inducing and Growth Inhibition Activities."J.Pesticide Sci.. (in press).
[Publications] H.Honda: "Induction of TNF-a gene expression by heat inducible promoter gadd"Japanese Journal of Hyperthermia Oncology.. 16(2). 91-98 (2000)
[Publications] H.Honda: "Heat induction of reporter gene expression via the gadd 153 promoter and its possible application to hyperthermia treatment of cancer."Cytotechnology.. 33. 131-137 (2000)
[Publications] H.Honda: "Bystander-killing effect and cyclic induction of TNF-a Gene under heat-inducible promoter gadd 153."Journal of Bioscience and Bioengineering.. 90(4). 437-441 (2000)
[Publications] H.Honda: "Preparation of tumor-specific magnetoliposomes and their application for hyperthermia."Journal of Chemical Engineering, Japan.. 34(1). 66-72 (2001)
[Publications] J.Ando: "Sp 1-mediated downregulation of P2X4 receptor gene transcription in endothelial cells exposed to shear stress."Am.J.Physiol Heart Circ.Physiol.. (in press).
[Publications] J.Ando: "mRNA expression on shape-engineered endothelial cells : Adhesion molecules ICAM-1 and VCAM-1."J.Biomed.Mater.Res.. 54. 366-372 (2001)
[Publications] J.Ando: "Fluid shear stress activates Ca^<2+> influx into human endothelial cells via P2X4 purinoceptors."Circ.Res.. 87. 385-391 (2000)
[Publications] J.Ando: "P2X4 receptors mediate ATP-induced calcium influx in human vascular endothelial cells."Am.J.Physiol Heart Circ.Physiol.. 279. 285-292 (2000)

2000 Fiscal Year Annual Research Report

単一細胞機能評価の新手法

Principal Investigator

松岡 英明 東京農工大学, 工学部, 教授 (10143653)

Research Products

[Publications] M.Saito: "pH microelectrode-micropipette system for the measurement of enzyme reactions in a picoliter volume of a living plant cell"Anal.Chim.Acta. 404(2). 223-229 (2000)

[Publications] M.Saito: "Elicitor action via cell membrane of a cultured rice cell demonstrated by the single-cell transient assay"J.Biotechnol.. 76(2/3). 227-232 (2000)

[Publications] M.Saito: "Modulation of Ca^<2+> influx into a single rice cell with an electric signal"Electrochemistry. 68(5). 333-336 (2000)

[Publications] M.Saito: "Use of acetylene for the fabrication of a glass capillary carbon microelec-trode"Electrochemistry. 68(11). 924-926 (2000)

[Publications] H.Matsuoka: "Microbioelectronics for single-cell experiment"Electrochemistry. 68(5). 314-320 (2000)

[Publications] 斉藤美佳子: "ボーダーレス電気化学者をめざす"電気化学. 69(2). 128-129 (2001)

[Publications] M.Natsume: "Effect of blue light on endogenous brassinosteroid activity and leaf unrolling in rice seedlings."ITE Letters on Batteroes, New Technologuies & Medicine. 1. 781-785 (2000)

[Publications] M.Natsume: "Teasterone-3-O-b-D-glucopyranoside, a new conjugated brassinosteroid metabolite from lily cell suspension cultures and its identification in lily anthors."Biosci.Biotech.Biochem.. 64. 702-709 (2000)

[Publications] M.Natsume: "Reversible Conversion between Teasterone and Its Ester Conjugates in Lily Cell Culture."J.Pesticide Sci.. 25. 117-122 (2000)

[Publications] M.Natsume: "Effects of De-N-methylation of Pamamycins on Aerial Mycelium-inducing and Growth Inhibition Activities."J.Pesticide Sci.. (in press).

[Publications] H.Honda: "Induction of TNF-a gene expression by heat inducible promoter gadd"Japanese Journal of Hyperthermia Oncology.. 16(2). 91-98 (2000)

[Publications] H.Honda: "Heat induction of reporter gene expression via the gadd 153 promoter and its possible application to hyperthermia treatment of cancer."Cytotechnology.. 33. 131-137 (2000)

[Publications] H.Honda: "Bystander-killing effect and cyclic induction of TNF-a Gene under heat-inducible promoter gadd 153."Journal of Bioscience and Bioengineering.. 90(4). 437-441 (2000)

[Publications] H.Honda: "Preparation of tumor-specific magnetoliposomes and their application for hyperthermia."Journal of Chemical Engineering, Japan.. 34(1). 66-72 (2001)

[Publications] J.Ando: "Sp 1-mediated downregulation of P2X4 receptor gene transcription in endothelial cells exposed to shear stress."Am.J.Physiol Heart Circ.Physiol.. (in press).

[Publications] J.Ando: "mRNA expression on shape-engineered endothelial cells : Adhesion molecules ICAM-1 and VCAM-1."J.Biomed.Mater.Res.. 54. 366-372 (2001)

[Publications] J.Ando: "Fluid shear stress activates Ca^<2+> influx into human endothelial cells via P2X4 purinoceptors."Circ.Res.. 87. 385-391 (2000)

[Publications] J.Ando: "P2X4 receptors mediate ATP-induced calcium influx in human vascular endothelial cells."Am.J.Physiol Heart Circ.Physiol.. 279. 285-292 (2000)

松岡英明東京農工大学, 工学部, 教授 (10143653)