2001 年度実績報告書

単一細胞機能評価の新手法

研究課題

研究課題/領域番号	11227202
研究機関	東京農工大学
研究代表者	松岡英明東京農工大学, 工学部, 教授 (10143653)
研究分担者	橋本壽正東京農工大学, 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)
キーワード	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+>
研究概要	本年度は各研究課題ごとに次のような成果を挙げた。 (1)単一細胞研究推進のためには,細胞集団から該当の細胞のみを選定し,掴み,移動し,所定の位置に置く,一連の操作の効率化を不可欠である。マルチウェルとマニピュレーションシステムおよびオートステージ,イメージスライサー型顕微分光システムを組み合わせることで,新たに「単一細胞遺伝子解析装置」を開発した。また,これを用いることで,イネ培養細胞に電気的刺激を印加し,Ca^<2+>の導入とそれに伴う,キチナーゼ遺伝子発現を観察することができた。これらの結果から,細胞内のCa^<2+>の増加が見られない場合には,キチナーゼ遺伝子が発現しないことが明らかになった。 (2)酵母細胞の電気化学的シグナルを印加した。この条件で生存率は20%まで減少した。このとき発現する遣伝子を遺伝子チップで解析した結果,非同調培養細胞を用いた場合には,TCA回路に関わる酵素やアセチルCoA代謝に関する酵素の発現抑制が認められたが,同調培養では同様な結果は確認されなかった。 (3)放線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。 (4)植物細胞中の熱拡散率解析を行った。顕微高速赤外線カメラを用いてペルチェ素子にて植物細胞を冷却したところ,水分が凝固するのは細胞単位で,発生はランダムで,かつ隣同士は同時に凝固しないことがわかった。 (5)熱シグナルに対するがん細胞の治癒条件の解析,流れせん断応力に対する内皮前駆細胞の反応になどを実施し,定量的な結果を得た。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。

研究成果
(26件)

すべてその他

すべて文献書誌 (26件)

[文献書誌] K.-B.Oh: "Purification and Characterization of an Autoregulatory substance Capable of regulating the morphological transition in Cnadida albicans"Proc.Natl.Acad.Sci. 98. 4664-4668 (2001)
[文献書誌] K.-B.Oh: "Detection of antifungal activity in Belamcanda chinensis by a Single-cell Bioassay method and isolation of its compound, Tectorigenin"Biosci.Biotechnol.Bioche.. 65. 939-942 (2001)
[文献書誌] H.Matsuoka: "Single-cell imaging of Ca2+ influx into bovine endotherial cells occurring in response to an alternating electric stimulus"Proceeding of Joint Symposium on Bio-Sensing and Bio-Imaging. (in press).
[文献書誌] M.Yokouchi: "Electrochemical killing of Streptococcus mutants causing dental caries"Electrochemistry. (in press).
[文献書誌] M.Natsume: "Phytotoxin production and werial mycelium formation by Streptomyces scabies and S. acidacabies in vitro"J.General Plant Pathol.. 67(in press). (2001)
[文献書誌] I.Kozone: "De-N-methylation of pamamycins on aerial mycelium indusing and growth inhibition activities"J.Pesticide Sci.. 26. 149-153 (2001)
[文献書誌] H.Honda: "Preparation of tumor-specific magnetoliposomes and their application for hyrperrtherima"J.Chem.Eng., Jpn. 34. 66-72 (2001)
[文献書誌] R.Korenaga: "Sp-1 mediated downregulation of P2X4 receptor gene transcription in endothelial cells exposed to Shear stress"Am.J.Physiol Heart Cric Physiol.. 280. 2214-2221 (2001)
[文献書誌] S.Kato: "MRNA expression on shape-engineered endothelial cells ; Adhesion molecules ICAM-1 and VCAM-1"J.Biomed Mater Res. 54. 366-372 (2001)
[文献書誌] H.Zhou: "Imaging localized horse radish peroxidase on glass sureface with scanning electrochemical/chemiluminescence microscopy"Anal.Biochem.. 290. 83-88 (2001)
[文献書誌] H.Shiku: "Oxygen consumption of single bovine embryos with scanning electrochemical microscopy"Anal Chem.. (in press).
[文献書誌] S.Ogata: "Dielectrophoretic manipulation of a single cell with dual-microdisk electrode"Bioelectrochemistry. (in press).
[文献書誌] M.nishiyama: "Substeps within the 8 nm step per ATPase cyclesof single kinesin molecules"Nat.Cell Biol.. (in press).
[文献書誌] T.Ito: "Determination of absolute concentration of nitroxide radial by radio-frequency EPR imaging"Appl.Magn.Reson.. 20. 257-263 (2001)
[文献書誌] G.Umitsuki: "Involvement of Rnase G in vivo mRNA metabolism in Escherichia coli"Genes Cells. 6. 403-410 (2001)
[文献書誌] M.Wachi: "Eacherichia coli ribonuclease G"Methods Enzymol.. 342. 55-63 (2001)
[文献書誌] K.kobayashi: "Suppression of the hypersensitivity of Escherichia coli acrB mutant to organic solvents by integrational activation of the acrEF operon with the IS1 or IS2 element"J.Bacteriol.. 183. 2646-2653 (2001)
[文献書誌] R.Ando: "Evaluation ofthe growth inhibition strength of hydrocarbon solvents against Escherichia coli and pseudomonas putida grown in a two-liquid phase culture system consisting of a medium and organic solvent"Extremophiles. 5. 11-15 (2001)
[文献書誌] A.Ito: "Heat-inducible TNF-a gene therapy combined with hyperthermia using magnetic nanoparticles as a novel tumortargeted therapy"Cancer gene therapy. 8(9). 649-654 (2001)
[文献書誌] S.Luo: "The dimerization of folded monomers of ribulose 1,5-bisphosphate carboxylase/oxtgenase"J.Biol.Chem. 276. 7023-7026 (2001)
[文献書誌] K.Nakasone: "Transcriptional regulation under pressure conditions by the RNA polymerase sigma54 factor with a two components regulatory system in Shewanella vilacea"Extremophiles. (in press).
[文献書誌] H.Matsuoka: "Simultaneous imaging of multiple fluorescent probes in bio-cells"Proceeding of Joint Symposium on Bio-Sensing and Bio-Imaging. (in press).
[文献書誌] 斉藤美佳子: "キャピラリーカーボンマイクロ電極(Galss capillary carbon microelectrode)"マテリアルインテグレーション. 15(2). 45-49 (2002)
[文献書誌] 松岡英明: "単一細胞実験技術の開発(Development of single-cell experiment technology)"バイオサイエンスとインダストリー.
[文献書誌] H.Matsuoka: "Single-cell viability assessment with a novel spectro-imaging"J.Biotechnol. 94(3). 299-308 (2002)
[文献書誌] 斉藤美佳子: "単一細胞実験法による遺伝子発現解析(Gene expression analysis by single-cell experiment)"生物物理. 41(5). 255-257 (2001)

2001 年度 実績報告書

単一細胞機能評価の新手法

研究代表者

松岡 英明 東京農工大学, 工学部, 教授 (10143653)

研究成果

[文献書誌] K.-B.Oh: "Purification and Characterization of an Autoregulatory substance Capable of regulating the morphological transition in Cnadida albicans"Proc.Natl.Acad.Sci. 98. 4664-4668 (2001)

[文献書誌] K.-B.Oh: "Detection of antifungal activity in Belamcanda chinensis by a Single-cell Bioassay method and isolation of its compound, Tectorigenin"Biosci.Biotechnol.Bioche.. 65. 939-942 (2001)

[文献書誌] H.Matsuoka: "Single-cell imaging of Ca2+ influx into bovine endotherial cells occurring in response to an alternating electric stimulus"Proceeding of Joint Symposium on Bio-Sensing and Bio-Imaging. (in press).

[文献書誌] M.Yokouchi: "Electrochemical killing of Streptococcus mutants causing dental caries"Electrochemistry. (in press).

[文献書誌] M.Natsume: "Phytotoxin production and werial mycelium formation by Streptomyces scabies and S. acidacabies in vitro"J.General Plant Pathol.. 67(in press). (2001)

[文献書誌] I.Kozone: "De-N-methylation of pamamycins on aerial mycelium indusing and growth inhibition activities"J.Pesticide Sci.. 26. 149-153 (2001)

[文献書誌] H.Honda: "Preparation of tumor-specific magnetoliposomes and their application for hyrperrtherima"J.Chem.Eng., Jpn. 34. 66-72 (2001)

[文献書誌] R.Korenaga: "Sp-1 mediated downregulation of P2X4 receptor gene transcription in endothelial cells exposed to Shear stress"Am.J.Physiol Heart Cric Physiol.. 280. 2214-2221 (2001)

[文献書誌] S.Kato: "MRNA expression on shape-engineered endothelial cells ; Adhesion molecules ICAM-1 and VCAM-1"J.Biomed Mater Res. 54. 366-372 (2001)

[文献書誌] H.Zhou: "Imaging localized horse radish peroxidase on glass sureface with scanning electrochemical/chemiluminescence microscopy"Anal.Biochem.. 290. 83-88 (2001)

[文献書誌] H.Shiku: "Oxygen consumption of single bovine embryos with scanning electrochemical microscopy"Anal Chem.. (in press).

[文献書誌] S.Ogata: "Dielectrophoretic manipulation of a single cell with dual-microdisk electrode"Bioelectrochemistry. (in press).

[文献書誌] M.nishiyama: "Substeps within the 8 nm step per ATPase cyclesof single kinesin molecules"Nat.Cell Biol.. (in press).

[文献書誌] T.Ito: "Determination of absolute concentration of nitroxide radial by radio-frequency EPR imaging"Appl.Magn.Reson.. 20. 257-263 (2001)

[文献書誌] G.Umitsuki: "Involvement of Rnase G in vivo mRNA metabolism in Escherichia coli"Genes Cells. 6. 403-410 (2001)

[文献書誌] M.Wachi: "Eacherichia coli ribonuclease G"Methods Enzymol.. 342. 55-63 (2001)

[文献書誌] K.kobayashi: "Suppression of the hypersensitivity of Escherichia coli acrB mutant to organic solvents by integrational activation of the acrEF operon with the IS1 or IS2 element"J.Bacteriol.. 183. 2646-2653 (2001)

[文献書誌] R.Ando: "Evaluation ofthe growth inhibition strength of hydrocarbon solvents against Escherichia coli and pseudomonas putida grown in a two-liquid phase culture system consisting of a medium and organic solvent"Extremophiles. 5. 11-15 (2001)

[文献書誌] A.Ito: "Heat-inducible TNF-a gene therapy combined with hyperthermia using magnetic nanoparticles as a novel tumortargeted therapy"Cancer gene therapy. 8(9). 649-654 (2001)

[文献書誌] S.Luo: "The dimerization of folded monomers of ribulose 1,5-bisphosphate carboxylase/oxtgenase"J.Biol.Chem. 276. 7023-7026 (2001)

[文献書誌] K.Nakasone: "Transcriptional regulation under pressure conditions by the RNA polymerase sigma54 factor with a two components regulatory system in Shewanella vilacea"Extremophiles. (in press).

[文献書誌] H.Matsuoka: "Simultaneous imaging of multiple fluorescent probes in bio-cells"Proceeding of Joint Symposium on Bio-Sensing and Bio-Imaging. (in press).

[文献書誌] 斉藤美佳子: "キャピラリーカーボンマイクロ電極(Galss capillary carbon microelectrode)"マテリアル インテグレーション. 15(2). 45-49 (2002)

[文献書誌] 松岡英明: "単一細胞実験技術の開発(Development of single-cell experiment technology)"バイオサイエンスとインダストリー.

[文献書誌] H.Matsuoka: "Single-cell viability assessment with a novel spectro-imaging"J.Biotechnol. 94(3). 299-308 (2002)

[文献書誌] 斉藤美佳子: "単一細胞実験法による遺伝子発現解析(Gene expression analysis by single-cell experiment)"生物物理. 41(5). 255-257 (2001)

2001 年度実績報告書

松岡英明東京農工大学, 工学部, 教授 (10143653)

[文献書誌] 斉藤美佳子: "キャピラリーカーボンマイクロ電極(Galss capillary carbon microelectrode)"マテリアルインテグレーション. 15(2). 45-49 (2002)