A new approach for the oocyte genotoxicity assay: Adaptation of comet assay on mouse cumulus–oocyte complexes

Abstract

Conventional genotoxicity tests are technically difficult to apply to oocytes, and results obtained on somatic cells cannot be extrapolated to gametes. We have previously described a comet assay (original–CA) on denuded mouse oocytes, but, in vivo, oocytes are not isolated from their surrounding follicular cells. Our objective was to develop a comet assay on cumulus–oocyte complexes (COC–CA) for a more physiological approach to study the genotoxicity of environmental factors on oocytes. For COC–CA, whole COC were exposed directly to exogenous agents after ovulation and removal from oviducts. Three conditions were studied: a negative control group, and two positive control groups, one of which was exposed to hydrogen peroxide (H2O2) and the other group was incubated with cerium dioxide nanoparticles (CeO2 NPs). With both tests, DNA damage was significant in the presence of both H2O2 and CeO2 NPs compared with the negative control. COC–CA offers an interesting tool for assaying the genotoxicity of environmental agents towards germinal cells. Furthermore, COC–CA is less timeconsuming and simplifies the protocol of the original–CA, because COC-CA is easier to perform without the washing-out procedure.