title: Embryonic Stage-Dependent Teratogenicity of Ketamine in Zebrafish (Danio rerio).
abstract: Ketamine, a widely used anesthetic, has been shown to have NMDA receptor dependent and independent actions during zebrafish (Danio rerio) embryogenesis. Notwithstanding, the effects of developmental toxicity and the mechanisms of ketamine action on fish embryos are still not well understood, and its implications for early vertebrate development remains to be clarified. In this work, zebrafish embryos were exposed to ketamine (0.2, 0.4, and 0.8 mg mL(-1)) in order to study the stage-developmental toxicity of this pharmaceutical. During 256-cell (2.5 h post-fertilization, hpf), 50% epiboly (5.5 hpf) and 1-4 somites (10.5 hpf), embryos were exposed to the referred ketamine concentrations for a period of 20 min and were allowed to grow until 144 hpf. Both lethal and nonlethal parameters were evaluated. Skeletal development was assessed by alcian blue and calcein staining. Additionally, the expression of the developmental genes sonic hedgehog a (shh a) and noggin 3 (nog3) was evaluated. Similar to our previous work, bone and cartilage malformations were observed after 256-cell exposure. During 50% epiboly, ketamine exposure induced concentration-dependent mortality and malformations, such as lordosis and/or kyphosis and microcephaly, namely, at higher concentrations. Conversely, exposure during 1-4 somites showed the induction of nonspecific effects with no rise in mortality. The quantitative real-time polymerase chain reaction (qRT-PCR) analysis showed differences in shh a and nog3 expressions comparatively to the control group. Overall, this study shows that the ketamine toxic profile is developmental phase-dependent with 256-cell being the most susceptible phase. The effects observed may result from ketamine interaction with cellular signaling pathways that merits further investigation.
authors.affiliation: Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Trás-os-Montes and Alto Douro (UTAD) , Vila Real, Portugal.|Laboratory Animal Science (LAS), Institute for Molecular and Cell Biology (IBMC), University of Porto (UP) , Porto, Portugal.|Institute for Research and Innovation in Health (i3S), University of Porto (UP) , Porto, Portugal.
authors.forename: Luís M
authors.initials: LM
authors.lastname: Félix
authors.affiliation: Life Sciences and Environment School (ECVA), University of Trás-os-Montes and Alto Douro (UTAD) , Vila Real, Portugal.
authors.forename: Cindy
authors.initials: C
authors.lastname: Serafim
authors.affiliation: Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Trás-os-Montes and Alto Douro (UTAD) , Vila Real, Portugal.|Laboratory Animal Science (LAS), Institute for Molecular and Cell Biology (IBMC), University of Porto (UP) , Porto, Portugal.|Institute for Research and Innovation in Health (i3S), University of Porto (UP) , Porto, Portugal.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Valentim
authors.affiliation: Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Trás-os-Montes and Alto Douro (UTAD) , Vila Real, Portugal.|Laboratory Animal Science (LAS), Institute for Molecular and Cell Biology (IBMC), University of Porto (UP) , Porto, Portugal.|Institute for Research and Innovation in Health (i3S), University of Porto (UP) , Porto, Portugal.|School of Agrarian and Veterinary Sciences (ECAV), University of Trás-os-Montes and Alto Douro (UTAD) , Vila Real, Portugal.
authors.forename: Luís M
authors.initials: LM
authors.lastname: Antunes
authors.affiliation: Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Trás-os-Montes and Alto Douro (UTAD) , Vila Real, Portugal.|Laboratory Animal Science (LAS), Institute for Molecular and Cell Biology (IBMC), University of Porto (UP) , Porto, Portugal.|Institute for Research and Innovation in Health (i3S), University of Porto (UP) , Porto, Portugal.
authors.forename: Sónia
authors.initials: S
authors.lastname: Campos
authors.affiliation: Biosystems & Integrative Sciences Institute (BioISI), Faculty of Sciences, University of Lisboa , Lisboa, Portugal.|Department of Genetics and Biotechnology (DGB), University of Trás-os-Montes and Alto Douro (UTAD) , Vila Real, Portugal.
authors.forename: Manuela
authors.initials: M
authors.lastname: Matos
authors.affiliation: Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Trás-os-Montes and Alto Douro (UTAD) , Vila Real, Portugal.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Coimbra
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D007649:Ketamine / Q000633:toxicity*; D013056:Spectrophotometry, Ultraviolet; D013723:Teratogens / Q000633:toxicity*; D015027:Zebrafish / Q000196:embryology*
chemical_list: D013723:Teratogens; D007649:Ketamine