title: Ex vivo assessment of testicular toxicity induced by carbendazim and iprodione, alone or in a mixture.
abstract: To measure the testicular toxicity of two fungicides (carbendazim and iprodione), alone or in a mixture, we used a rat ex vivo model of seminiferous tubules, greatly reducing the number of rodents used, in accordance with the 3R rule (Replacement, Reduction, and Refinement). This model allows the representation of puberty, a critical life period with regard to endocrine disruptors. The cellular modifications were followed for three weeks through transcriptomic and proteomic profiling analysis. A quantitative and comparative method was developed to estimate how known pathways were disturbed by each substance. This pathway-driven analysis revealed a strong alteration of steroidogenesis and an impairment of meiosis in all cases, albeit the initial molecular events were different for both substances. The ex vivo cytogenetic analysis confirmed that both fungicides alter the course of the first meiotic prophase. In addition, the mixture of both substances triggered effects greater than the sum of their cumulative effects and compromised future sperm motility after a shorter time of exposure compared with the fungicides tested separately. The alliance of an ex vivo culture with "omics" strategies complemented with a physiological examination is a powerful combination of tools for testing substances, separately or in a mixture, for their testicular toxicity. In particular, proteomics allowed the identification of systematically differentially expressed proteins in the secretomes of exposed cultures, such as FUCO and PEBP1, two proteins linked with the motility and fertilizing ability of spermatozoa, respectively. These proteins may be potential biomarkers of testicular dysfunction and infertility.
authors.affiliation: CEA, Direction de la Recherche Fondamentale, BIAM-Marcoule, Bagnols-sur-Cèze, France.
authors.forename: Cédric
authors.initials: C
authors.lastname: Pisani
authors.affiliation: Plateforme BioPark d'Archamps, Archamps Technopole, Saint Julien en Genevois, France.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Voisin
authors.affiliation: Plateforme BioPark d'Archamps, Archamps Technopole, Saint Julien en Genevois, France.
authors.forename: Karim
authors.initials: K
authors.lastname: Arafah
authors.affiliation: IGFL, UMR 5242 CNRS INRA Ecole Normale Supérieure de Lyon, Lyon, France.|Current address: Kallistem, SAS, ENS, Lyon, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Durand
authors.affiliation: INSERM U1208, Stem Cell and Brain Research Institute, Université Lyon 1, Bron, France.
authors.forename: Marie-Hélène
authors.initials: MH
authors.lastname: Perrard
authors.affiliation: IMBE, UMR CNRS 7263 / IRD 237, Aix-Marseille Université (AMU), Marseille, France.
authors.forename: Marie-Roberte
authors.initials: MR
authors.lastname: Guichaoua
authors.affiliation: Plateforme BioPark d'Archamps, Archamps Technopole, Saint Julien en Genevois, France.|Université Grenoble Alpes, Institut Albert Bonniot, Inserm 1209, CNRS UMR5309, Equipe Immunologie Analytique des Pathologies Chroniques, Grenoble, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Bulet
authors.affiliation: CEA, Direction de la Recherche Fondamentale, BIAM-Marcoule, Bagnols-sur-Cèze, France.
authors.forename: Odette
authors.initials: O
authors.lastname: Prat
mesh_terms: D000620:Aminoimidazole Carboxamide / Q000031:analogs & derivatives* / Q000633:toxicity; D000826:Animal Testing Alternatives / Q000379:methods*; D000818:Animals; D001562:Benzimidazoles / Q000633:toxicity*; D002219:Carbamates / Q000633:toxicity*; D005659:Fungicides, Industrial / Q000633:toxicity; D006827:Hydantoins / Q000633:toxicity*; D008297:Male; D008540:Meiosis / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D012671:Seminiferous Tubules / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D012729:Sex Chromosome Aberrations / Q000187:drug effects; D013091:Spermatogenesis / Q000187:drug effects; D013256:Steroids / Q000096:biosynthesis; D013733:Testicular Diseases / Q000139:chemically induced*; D046509:Tissue Culture Techniques; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D001562:Benzimidazoles; D002219:Carbamates; D005659:Fungicides, Industrial; D006827:Hydantoins; D013256:Steroids; D000620:Aminoimidazole Carboxamide; C006698:carbendazim; C033148:iprodione
keywords: biomarker; endocrine disruption; pesticides; spermatogenesis; toxicogenomics