Bio-Cell Gene Expression Analyzer
Bio-Cell Gene Expression Analyzer
批准号:
09355027
负责人:
MATSUOKA Hideaki
金额:
$19.2万
依托单位国家:
日本
项目类别:
Grant-in-Aid for Scientific Research (A).
财政年份:
1997
资助国家:
日本
项目状态:
已结题
起止时间:
1997 至 2000
中文摘要
术语“生物细胞”指的是在胁迫作用的整个实验过程中保持其存活状态的细胞。这项研究是为了演示生物细胞对各种应激信号的反应的实时分析。电应力信号被施加到烟草或水稻的培养细胞上。作为细胞反应,用显微荧光图像分析系统测量基因表达。通过烟草细胞株BY-2的实验,测量了不同强度的脉冲电应力信号下细胞膜的电阻抗。结果表明,当跨膜电位(V<;Cp>;)超过200 mV时,膜有明显的通透性。另一方面,利用水稻原生质体进行的两步实验证明,几丁质酶基因的表达依赖于钙离子。在第一步,由脉冲电应力信号控制钙离子的引入。结果表明,在VCMP的1-5 mV范围内,Ca~(2+)&Gt~(2+)进入细胞内。作为第二步,我们将Ca~(2+)&Gt~(2+)直接注入原生质体中,证实了几丁质酶基因的表达。除了电应力,我们还打算在生物细胞内部施加电化学应力。然后,我们研制了直径为1-2μm的毛细碳微电极,并通过分析多巴胺的循环伏安图来检测其性能。综上所述,开发了生物细胞基因表达分析系统,并展示了其良好的性能。
英文摘要
The term "bio-cell" means a cell that maintains its viable state throughout the experiments of stress application. This study was done in order to demonstrate the real time analysis of the bio-cell responses to various stress signals. An electric stress signal was applied to cultured cells of tobacco or rice. As the cell response, the gene expression was measured with a microscopic fluorescent image analyzing system. By the experiment using tobacco cells, line BY-2, the electric impedance of the cell membrane was measured under various intensities of pulsing electric stress signals. As the result, appreciable membrane permeabilization was observed when the cross membrane potential (V_<CMP>) exceeded 200mV.On the other hand, the Ca^<2+>-dependent expression of chitinase gene was demonstrated by 2-step experiments using rice protoplasts. At the first step, Ca^<2+> introduction was controlled by the pulsing electric stress signal. As the result, Ca^<2+> entered the cells around 1-5 mV of VCMP.As the 2nd step, we demonstrated the chitinase gene expression by the direct injection of Ca^<2+> into the protoplasts. Besides the electric stress, we intended to apply an electrochemical stress to a bio-cell at its inside. Then we developed a capillary carbon microelectrode with a 1-2 μm diameter and checked its performance by analyzing cyclicvoltammograms of dopamine. In conclusion, the analyzing system of the gene expression of a bio-cell was developed and its promising performance was demonstrated.
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Mikako Saito, Ayako Saga, Hideaki Matsuoka and Danny K.Y.Wong: "Use of acetylene for the fabrication of a glass capillary carbon microelectrode"Electrochemistry. 68(11). 924-926 (2000)
Mikako Saito、Ayako Saga、Hideaki Matsuoka 和 Danny K.Y.Wong:“使用乙炔制造玻璃毛细管碳微电极”电化学。
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Hidekazu Sotoyama: "Control of the Cross Membrane Potential of a Cultured Tobacco Cell and Simultaneous Measurement of the Electrical Impedance of Its Membrane." Electrochemistry. 67(1). 18-21 (1999)
Hidekazu Sotoyama:“控制培养烟草细胞的跨膜电位并同时测量其膜的电阻抗。”
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Hideaki Matsuoka: "Use of a Microelectrode System as a Tool of the Quantitative Application of Wound to the Cell-to-Cell Junctional Membrane of Cultured Tobacco Cells." Denki Kagaku. (in press). (1998)
Hideaki Matsuoka:“使用微电极系统作为定量应用伤口到培养的烟草细胞的细胞间连接膜的工具。”
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Hideaki Matsuoka: "Effects of a pulsing electric signal on the Cross Membrane Potential and the Cell Division Potentiality of a Single Cell of Tabacco."Bioelectrochem. Bioenerg.. 49. 65-72 (1999)
Hideaki Matsuoka:“脉冲电信号对烟草单细胞跨膜电位和细胞分裂电位的影响。”生物电化学。
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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)
Mikako Saito:“致力于成为无国界电化学家”电化学 69(2)。
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共 38 条
Analysis of dynamic control mechanism of undifferentiated state in ES cells
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批准号:23246142
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项目类别:Grant-in-Aid for Scientific Research (A)
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资助金额:$31.12万
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财政年份:2011
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负责人:MATSUOKA Hideaki
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依托单位:
Supervision of the Research of Single-cell Molecular Technology
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批准号:11227101
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项目类别:Grant-in-Aid for Scientific Research on Priority Areas
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资助金额:$16.77万
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财政年份:1999
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负责人:MATSUOKA Hideaki
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依托单位:
Rapid Bioassay with a Bio-Cell Tracer System (BCT)
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批准号:07044131
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项目类别:Grant-in-Aid for international Scientific Research
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资助金额:$9.22万
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财政年份:1995
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负责人:MATSUOKA Hideaki
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依托单位:
Development of an Analyzing System of the Cellular Responses to Multiple Stresses
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批准号:06555245
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项目类别:Grant-in-Aid for Scientific Research (A)
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资助金额:$8.38万
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财政年份:1994
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负责人:MATSUOKA Hideaki
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依托单位:
Analysis of Intercellular Interaction in a Process of Neurite Growth
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批准号:04454617
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项目类别:Grant-in-Aid for General Scientific Research (B)
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资助金额:$3.9万
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财政年份:1992
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负责人:MATSUOKA Hideaki
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依托单位:
Development of Immunoanalyzer Based on Gate Function of Protein Membrane
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批准号:60850141
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项目类别:Grant-in-Aid for Developmental Scientific Research
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资助金额:$3.2万
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财政年份:1985
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负责人:MATSUOKA Hideaki
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依托单位:
海外基金