Live Imaging of Radiation-Induced Apoptosis by Yolk Injection of Acridine Orange in the Developing Optic Tectum of Medaka
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概要
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To observe the sequential radiation-induced apoptosis in a living embryo, we injected Acridine Orange (AO) solution into the yolk of embryo and visualized radiation-induced apoptosis in developing optic tectum (OT). Medaka embryos at stage 28, when neural cells proliferate rapidly in the OT, were irradiated with 5 Gy X-rays which is a non-lethal dose for irradiated embryos at hatching. The irradiated embryos hatched normally without morphological abnormalities in their brains, even though a large number of apoptotic cells were induced transiently in OT. By yolk injection, apoptotic cells in OT were distinguished as AO-positive small nuclei at 3 h after irradiation. At 8-10 h after irradiation, AO-positive rosette-shaped clusters were obviously distinguished in marginal tectal regions of OT where cells are proliferating intensely. The AO-positive clusters became bigger and more obvious, but the number did not increase up to 24 h after irradiation and completely disappeared up to 49 h after irradiation. This characteristic appearance of the AO-positive nuclei/clusters is in good agreement with our previous results, based on the examination of fixed specimens stained with AO by injection into the peri-vitelline space, suggesting that the AO-yolk injection method is highly reliable for detecting apoptotic cells in living embryos. The live imaging of apoptotic cells in developing Medaka embryos by AO-yolk injection method is expected to reveal more of the details of the dynamics of apoptotic responses in the irradiated brain and other tissues.
- 2009-11-16
著者
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YASUDA Takako
Research Center for Charged Particle Therapy, National Institute of Radiological Sciences
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ODA Shoji
Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo
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ISHIKAWA Yuji
Research Center for Radiation Protection, National Institute of Radiological Sciences
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WATANABE ASAKA
Space Biomedical Research Office, Japan Aerospace Exploration Agency
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HIDAKA Masayuki
Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo
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YASUDA Hiroshi
Research Center for Radiation Protection, National Institute of Radiological Sciences
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ANZAI Kazunori
Research Center for Charged Particle Therapy, National Institute of Radiological Sciences
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MITANI Hiroshi
Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo
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Mitani Hiroshi
Department Of Integrated Biosciences Graduate School Of Frontier Sciences The University Of Tokyo
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Anzai Kazunori
Research Center For Charged Particle Therapy National Institute Of Radiological Sciences
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Yasuda Takako
Research Center For Charged Particle Therapy National Institute Of Radiological Sciences
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Watanabe Asaka
Space Biomedical Research Office Japan Aerospace Exploration Agency
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Oda Shoji
Department Of Integrated Biosciences Graduate School Of Frontier Sciences The University Of Tokyo
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Ishikawa Yuji
Research Center For Radiation Protection National Institute Of Radiological Sciences
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Hidaka Masayuki
Department Of Integrated Biosciences Graduate School Of Frontier Sciences The University Of Tokyo
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Mitani Hiroshi
Department Of Biological Sciences School Of Science University Of Tokyo.
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Yasuda Hiroshi
Research Center For Advanced Science And Technology University Of Tokyo
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Yasuda Hiroshi
Research Center For Radiation Protection National Institute Of Radiological Sciences
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Yasuda Hiroshi
Research Center For Advanced Science And Technology The University Of Tokyo
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Mitani Hiroshi
Department Of Biological Sciences Graduate School Of Frontier Sciences University Of Tokyo
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