1999 Fiscal Year Final Research Report Summary
Development of method for detection of DNA damages by the comet assay with Daphnia magna
| Project/Area Number |
10680525
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
環境影響評価(含放射線生物学)
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| Research Institution | Tokyo University of Pharmacy and Life Science |
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Principal Investigator |
OHTA Toshihiro Tokyo University of Pharmacy and Life Science, School of Life Science, Associate professor, 生命科学部, 助教授 (10266893)
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| Project Period (FY) |
1998 – 1999
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| Keywords | Daphnia magna / Comet assay / SCG assay / DNA damage / Mutagen / Alkylating agent |
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| Research Abstract |
A method to detect DNA damages in vivo induced by environmental mutagens in water flea, Daphnia magna, was investigated. Recent development of the in vivo alkaline single cell gel electrophoresis assay (SCG assay, comet assay) facilitates the detection of in vivo genotoxicity in multiple organs from the same animal. We modified the new comet assay method for the nuclei of D.magna cells. Conditions to obtain nuclei fraction from the homogenate of D.magna cells were determined. When D.magna was treated with methyl methane sulfonate (MMS) at 50 μg/ml or more for 30 min, the frequency of DNA damaged nuclei was apparently higher compared with that observed in untreated control. It is considered that much improvement in extraction and purification of nuclei from homogenate is needed.
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