COMBINATION OF FEULGEN NUCLEAR REACTION WITH IMMUNOFLUORESCENT STAINING FOR PHOTOPRODUCTS OF DNA AFTER UV-IRRADIATION
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概要
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Immunofluorescent staining for nuclear UV photoproducts was combined with nuclear Feulgen reaction for cytofluorometry to quantitate the potency of excision repair of the lesions per unit amount of DNA on a single cell. Isolated hepatocytes and cerebellar neurons of adult rat were UV-irradiated and made react with rabbit antiserum containing antibodies to photoproducts of DNA after UV-irradiation. The UV induced lesions were demonstrated by indirect immunofluorescent technique using FITC labeled anti-rabbit's immunoglobulin antibody. The immunofluorescent staining was combined with subsequent Feulgen nuclear reaction after stabilizing the fluorescent dye-immunoglubulin complex by post-fixation with absolute ethanol. As a linear relationship was found between fluorescence intensity and concentration of free and protein-bound FITC up to absorbance value of 0.1, intensity of the immunofluorescent staining was controled not to exceed this limit. Two peaks of fluorescence of FITC (520 nm) and Feulgen dye (613 nm) were separately identified on a double-stained nucleus with respective excitation by 450 nm and 543 nm without interference of each other fluorescence. Proportionality between DNA content and amount of Feulgen fluorescence was preserved in the double stained hepatocytes and cerebellar neurons. Linear relationships were noted between the amounts of Feulgen DNA and nuclear FITC in the same cells immediately after UV-irradiation. The results indicate that the amounts of photoproducts produced in unit amount of nuclear DNA are equal in all cells irrespective of the kinds of tissues and stages of cell cycle.
- 日本組織細胞化学会の論文
著者
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Shima Akihiro
Laboratory of Radiation Biology, Department of Biological Sciences, School of Science, University of
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FUKUDA Masaru
Department of Pathology, Fukui Medical University
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Shima Akihiro
Laboratory Of Radiation Biology Faculty Of Science University Of Tokyo
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NAKANISHI KAZUO
Department of Pathology, Kyoto Prefectural University of Medicine
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FUJITA SETSUYA
Department of Pathology, Kyoto Prefectural University of Medicine
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Fujita Setsuya
Department Of Hygiene Gifu University School Of Medicine
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Fujita Setsuya
Dept. Of Ophthalmol. And Pathol. Kyoto Pref. Univ. Med.
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Fujita Setsuya
Louis Pasteur Center For Medical Research
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MUKAINAKA Teruo
Kyoto Prefectural University of Medicine
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MUKAINAKA Teruo
Department of Biochemistry, Kyoto Prefectural University of Medicine
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Fukuda Masaru
Department Of Pathology Kyoto Prefectural University Of Medicine
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Fukuda Masaru
Department Of Pathology Fukui Medical School
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Mukainaka Teruo
Department Of Pathology Kyoto Prefectural University Of Medicine
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Nakanishi K
Food Research And Development Laboratories Ajinomoto Co. Inc.
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Nakanishi Kazuo
Department Of Orthopaedic Surgery Okayama University Hospital Of Medicine And Dentistry
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Nakanishi Kazuo
Dept Of Pathology Kyoto Pref. Univ. Of Med.
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Shima Akihiro
Laboratory Of Radiation Biology Department Of Biological Sciences The University Of Tokyo
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Fujita Setsuya
Department Of Dynamic Pathology Research Institute For Neurological Diseases And Geriatrics Kyoto Pr
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Nakanishi Kazuo
Department Of Anesthesiology And Resuscitology Ehime University Graduate School Of Medicine
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Fukuda Masaru
Department Of Anatomy Kyorin University School Of Medicine
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Muranaka Takashi
Faculty Of Pharmaceutical Sciences Okayama University
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FUKUDA MASARU
Department of Pathology, Kyoto Prefectural University of Medicine
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