title: 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 time-consuming and simplifies the protocol of the original-CA, because COC-CA is easier to perform without the washing-out procedure.
authors.affiliation: Aix Marseille Université, CNRS, IRD, Avignon Université, IMBE UMR 7263, Marseille, France.
authors.forename: F
authors.initials: F
authors.lastname: Greco
authors.affiliation: Aix Marseille Université, CNRS, IRD, Avignon Université, IMBE UMR 7263, Marseille, France CEREGE, Aix-Marseille Université, CNRS, UM34, UMR 7330, Aix en Provence, France CECOS - Laboratory of Reproductive Medicine, AP-HM La Conception, Marseille, France.
authors.forename: J
authors.initials: J
authors.lastname: Perrin
authors.affiliation: CEREGE, Aix-Marseille Université, CNRS, UM34, UMR 7330, Aix en Provence, France.
authors.forename: M
authors.initials: M
authors.lastname: Auffan
authors.affiliation: Aix Marseille Université, CNRS, IRD, Avignon Université, IMBE UMR 7263, Marseille, France.
authors.forename: V
authors.initials: V
authors.lastname: Tassistro
authors.affiliation: Aix Marseille Université, CNRS, IRD, Avignon Université, IMBE UMR 7263, Marseille, France.
authors.forename: T
authors.initials: T
authors.lastname: Orsière
authors.affiliation: Aix Marseille Université, CNRS, IRD, Avignon Université, IMBE UMR 7263, Marseille, France Department of Gynaecology, Obstetrics, and Reproduction, AP-HM La Conception, Marseille, France blandine.courbiere@imbe.fr blandine.courbiere@ap-hm.fr.
authors.forename: B
authors.initials: B
authors.lastname: Courbiere
mesh_terms: D000818:Animals; D002563:Cerium / Q000633:toxicity; D020552:Comet Assay / Q000379:methods*; D054885:Cumulus Cells / Q000187:drug effects*; D004249:DNA Damage / Q000187:drug effects; D005260:Female; D006861:Hydrogen Peroxide / Q000633:toxicity; D053768:Metal Nanoparticles / Q000633:toxicity; D051379:Mice; D009153:Mutagens / Q000633:toxicity*; D009865:Oocytes / Q000187:drug effects*
chemical_list: D009153:Mutagens; D002563:Cerium; C030583:ceric oxide; D006861:Hydrogen Peroxide
keywords: comet assay; cumulus–oocyte complexes; environment; genotoxicity; oocyte