title: Visualizing Compound Distribution during Zebrafish Embryo Development: The Effects of Lipophilicity and DMSO.
abstract: The predictability of the zebrafish embryo model is highly influenced by internal exposure of the embryo/larva. As compound uptake is likely to be influenced by factors such as lipophilicity, solvent use, and chorion presence, this article focuses on investigating their effects on compound distribution within the zebrafish embryo. To visualize compound uptake and distribution, zebrafish embryos were exposed for 96 hr, starting at 4 hr postfertilization, to water-soluble dyes: Schiff's reagent (logP -4.63), Giemsa stain (logP -0.77), Van Gierson stain (logP 1.64), Cresyl fast violet (logP 3.5), Eosine Y (logP 4.8), Sudan III (logP 7.5), and Oil red O (logP 9.81), with and without 1% dimethyl-sulfoxide (DMSO). Three additional compounds were used to analytically determine the uptake and distribution: Acyclovir (logP -1.56), Zidovudine (logP 0.05), and Metoprolol Tartrate Salt (logP 1.8). Examinations were performed every 24 hr. Both methods (visualization and specific analysis) showed that exposure to higher logP values results in higher compound uptake. Specific analysis showed that for lipophilic compounds >90% of compound is taken up by the embryo. For hydrophilic compounds, >90% of compound within the complete egg could not be associated to embryo or chorion and is probably distributed into the perivitelline space. Overall, internal exposure analyses on at least two occasions (i.e., before and after hatching) is crucial for interpretation of zebrafish embryotoxicity data, especially for compounds with extreme logP values. DMSO did not affect exposure when examined with the visualization method, however, this method might be not sensitive enough to draw hard conclusions.
authors.affiliation: WIL Research Europe B.V, Hambakenwetering 7, 's-Hertogenbosch, The Netherlands.
authors.forename: Coco
authors.initials: C
authors.lastname: de Koning
authors.affiliation: WIL Research Europe B.V, Hambakenwetering 7, 's-Hertogenbosch, The Netherlands.
authors.forename: Manon
authors.initials: M
authors.lastname: Beekhuijzen
authors.affiliation: WIL Research Europe B.V, Hambakenwetering 7, 's-Hertogenbosch, The Netherlands.
authors.forename: Marysia
authors.initials: M
authors.lastname: Tobor-Kapłon
authors.affiliation: WIL Research Europe B.V, Hambakenwetering 7, 's-Hertogenbosch, The Netherlands.
authors.forename: Selinda
authors.initials: S
authors.lastname: de Vries-Buitenweg
authors.affiliation: WIL Research Europe B.V, Hambakenwetering 7, 's-Hertogenbosch, The Netherlands.
authors.forename: Dick
authors.initials: D
authors.lastname: Schoutsen
authors.affiliation: WIL Research Europe B.V, Hambakenwetering 7, 's-Hertogenbosch, The Netherlands.
authors.forename: Nico
authors.initials: N
authors.lastname: Leeijen
authors.affiliation: WIL Research Europe B.V, Hambakenwetering 7, 's-Hertogenbosch, The Netherlands.
authors.forename: Beppy
authors.initials: B
authors.lastname: van de Waart
authors.affiliation: WIL Research Europe B.V, Hambakenwetering 7, 's-Hertogenbosch, The Netherlands.
authors.forename: Harry
authors.initials: H
authors.lastname: Emmen
mesh_terms: D000212:Acyclovir / Q000494:pharmacology; D000818:Animals; D002853:Chromatography, Liquid; D004396:Coloring Agents / Q000378:metabolism*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000378:metabolism*; D047108:Embryonic Development / Q000187:drug effects*; D007814:Larva / Q000187:drug effects; D008055:Lipids / Q000494:pharmacology*; D013058:Mass Spectrometry; D008790:Metoprolol / Q000494:pharmacology; D010063:Ovum / Q000187:drug effects / Q000378:metabolism; D012996:Solutions; D015027:Zebrafish / Q000196:embryology*; D015215:Zidovudine / Q000494:pharmacology
chemical_list: D004396:Coloring Agents; D008055:Lipids; D012996:Solutions; D015215:Zidovudine; D008790:Metoprolol; D000212:Acyclovir; D004121:Dimethyl Sulfoxide
keywords: 3R's; compound uptake; dimethyl-sulfoxide (DMSO); internal exposure; lipophilicity; logP; solvent; zebrafish embryos (Danio rerio)