title: Chronic effects of clofibric acid in zebrafish (Danio rerio): a multigenerational study.
abstract: Clofibric acid (CA) is an active metabolite of the blood lipid lowering agent clofibrate, a pharmaceutical designed to work as agonist of peroxisome proliferator-activated receptor alpha (PPARa). It is the most commonly reported fibrate in aquatic environments with low degradation rate and potential environmental persistence. Previous fish exposures showed that CA may impact spermatogenesis, growth and the expression of fat binding protein genes. However, there are limited data on the effects of chronic multigenerational CA exposures. Here, we assessed chronic multigenerational effects of CA exposure using zebrafish (Danio rerio) as a teleost model. Zebrafish were exposed through the diet to CA (1 and 10mg/g) during their whole lifetime. Growth, reproduction-related parameters and embryonic development were assessed in the exposed fish (F1 generation) and their offspring (F2 generation), together with muscle triglyceride content and gonad histology. In order to study the potential underlying mechanisms, the transcription levels of genes coding for enzymes involved in lipid metabolism pathways were determined. The results show that chronic life-cycle exposure to CA induced a significant reduction in growth of F1 generation and lowered triglyceride muscle content (10mg/g group). Also, an impact in male gonad development was observed together with a decrease in the fecundity (10mg/g group) and higher frequency of embryo abnormalities in the offspring of fish exposed to the lowest CA dose. The profile of the target genes was sex- and tissue-dependent. In F1 an up-regulation of male hepatic pparaa, pparb and acox transcript levels was observed, suggesting an activation of the fatty acid metabolism (provided that transcript level change indicates also a protein level change). Interestingly, the F2 generation, raised with control diet, displayed a response pattern different from that observed in F1, showing an increase in weight in the descendants of CA exposed fish, in comparison with control animals, which points to a multigenerational effect.
authors.affiliation: Centre for The Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Trás-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5000-801 Vila Real, Portugal; Department of Biology and Environment, Life Sciences and Environment School (ECVA), University of Trás-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5000-801 Vila Real, Portugal. Electronic address: acoimbra@utad.pt.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Coimbra
authors.affiliation: CIMAR/CIIMAR, Interdisciplinary Centre for Marine and Environmental Research, University of Porto, Rua dos Bragas 177, 4050-123 Porto, Portugal; Department of Biology and Environment, Life Sciences and Environment School (ECVA), University of Trás-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5000-801 Vila Real, Portugal.
authors.forename: Maria João
authors.initials: MJ
authors.lastname: Peixoto
authors.affiliation: CIMAR/CIIMAR, Interdisciplinary Centre for Marine and Environmental Research, University of Porto, Rua dos Bragas 177, 4050-123 Porto, Portugal.
authors.forename: Inês
authors.initials: I
authors.lastname: Coelho
authors.affiliation: CIMAR/CIIMAR, Interdisciplinary Centre for Marine and Environmental Research, University of Porto, Rua dos Bragas 177, 4050-123 Porto, Portugal.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Lacerda
authors.affiliation: CIMAR/CIIMAR, Interdisciplinary Centre for Marine and Environmental Research, University of Porto, Rua dos Bragas 177, 4050-123 Porto, Portugal; FCUP, Faculty of Sciences University of Porto, Department of Biology, Rua do Campo Alegre, 4169-007 Porto, Portugal.
authors.forename: António Paulo
authors.initials: AP
authors.lastname: Carvalho
authors.affiliation: CIMAR/CIIMAR, Interdisciplinary Centre for Marine and Environmental Research, University of Porto, Rua dos Bragas 177, 4050-123 Porto, Portugal; Department of Functional Biology and Health Sciences, Faculty of Biology, University of Vigo, As Lagoas-Marcosende s/n, 36310, Vigo, Spain.
authors.forename: Manuel
authors.initials: M
authors.lastname: Gesto
authors.affiliation: CIMAR/CIIMAR, Interdisciplinary Centre for Marine and Environmental Research, University of Porto, Rua dos Bragas 177, 4050-123 Porto, Portugal.
authors.forename: Angeliki
authors.initials: A
authors.lastname: Lyssimachou
authors.affiliation: CIMAR/CIIMAR, Interdisciplinary Centre for Marine and Environmental Research, University of Porto, Rua dos Bragas 177, 4050-123 Porto, Portugal.
authors.forename: Daniela
authors.initials: D
authors.lastname: Lima
authors.affiliation: CIMAR/CIIMAR, Interdisciplinary Centre for Marine and Environmental Research, University of Porto, Rua dos Bragas 177, 4050-123 Porto, Portugal.
authors.forename: Joana
authors.initials: J
authors.lastname: Soares
authors.affiliation: CIMAR/CIIMAR, Interdisciplinary Centre for Marine and Environmental Research, University of Porto, Rua dos Bragas 177, 4050-123 Porto, Portugal.
authors.forename: Ana
authors.initials: A
authors.lastname: André
authors.affiliation: CIMAR/CIIMAR, Interdisciplinary Centre for Marine and Environmental Research, University of Porto, Rua dos Bragas 177, 4050-123 Porto, Portugal.
authors.forename: Ana
authors.initials: A
authors.lastname: Capitão
authors.affiliation: CIMAR/CIIMAR, Interdisciplinary Centre for Marine and Environmental Research, University of Porto, Rua dos Bragas 177, 4050-123 Porto, Portugal; FCUP, Faculty of Sciences University of Porto, Department of Biology, Rua do Campo Alegre, 4169-007 Porto, Portugal.
authors.forename: Luís Filipe C
authors.initials: LF
authors.lastname: Castro
authors.affiliation: CIMAR/CIIMAR, Interdisciplinary Centre for Marine and Environmental Research, University of Porto, Rua dos Bragas 177, 4050-123 Porto, Portugal; FCUP, Faculty of Sciences University of Porto, Department of Biology, Rua do Campo Alegre, 4169-007 Porto, Portugal. Electronic address: santos@ciimar.up.pt.
authors.forename: Miguel M
authors.initials: MM
authors.lastname: Santos
mesh_terms: D000818:Animals; D001835:Body Weight / Q000187:drug effects; D002995:Clofibric Acid / Q000633:toxicity*; D047108:Embryonic Development / Q000187:drug effects; D005298:Fertility / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D006066:Gonads / Q000187:drug effects; D000960:Hypolipidemic Agents / Q000633:toxicity; D008297:Male; D012744:Sex Ratio; D014874:Water Pollutants, Chemical / Q000633:toxicity; D015027:Zebrafish / Q000502:physiology*
chemical_list: D000960:Hypolipidemic Agents; D014874:Water Pollutants, Chemical; D002995:Clofibric Acid
keywords: Emerging pollutant; Endocrine disruption; Epigenetic; Lipid metabolism; Multigenerational; Reproduction