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Use of Frozen Tissue in the Comet Assay for the Evaluation of DNA Damage
Published on: March 24, 2020
Linking genotoxic responses in Gammarus fossarum germ cells with reproduction impairment, using the Comet assay
Emilie Lacaze1, Olivier Geffard, Delphine Goyet
1Université de Lyon, INRA-ENTPE, Laboratoire des Sciences de l'Environnement, rue Maurice Audin, Vaulx en Velin, F-69518, France. emilie.lacaze@cemagref.fr
Abstract:
Germ cells perform a unique and critical biological function: they pass down DNA that will be used for the development of the next generation. Thus there is an increasing need to understand how the adult exposure to genotoxicants could show negative impact on the offspring of aquatic organisms. Hence this work addresses the question of the consequences of germ cell DNA damage resulting from parental exposure on reproduction quality in the freshwater crustacean Gammarus fossarum, a high ecologically relevant species. Initially, the sensitivity response of mature oocytes and spermatozoa to two model genotoxicants, MMS and K(2)Cr(2)O(7) was compared by implementing the Comet assay after the exposure of these gammarids in the laboratory and after the exposure of caged organisms in the field. Spermatozoa appeared significantly more susceptible than the oocytes to genotoxicants whatever were the exposure conditions. Secondly, a significant correlation between the level of damage to the sperm DNA of exposed parents and the abnormality rate in embryos that had developed in non-contaminated water were demonstrated. Interestingly, this relationship bridges the biomarker response measured in germ cells at molecular level and its consequences at individual level for the subsequent generation. Moreover, reproduction defects were observed for a level of DNA damage exceeding a minimal threshold, which could have significant consequences for the population dynamics of this high ecologically relevant species.
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