| Co-Investigator(Kenkyū-buntansha) |
SAITO Mikako Tokyo Univ. Agricul. Technol., Dept. Technol., Research Assistant, 工学部, 講師 (20291346)
OH Ki-bong Tokyo Univ. Agricul. Technol., Dept. Technol., Assistant Professor (90272632)
NEMOTO Yasuyuki Tokyo Univ. Agricul. Technol., Dept. Technol., Assistant Professor (70202249)
KAWANA Akio NTT Basic Research Laboratories, Material Science Research Laboratory, Kawana Research Group Leader
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| Research Abstract |
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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