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Visualizing Compound Distribution During Zebrafish Embryo Development: The Effects of Lipophilicity and DMSO

Visualizing Compound Distribution During Zebrafish Embryo Development: The Effects of Lipophilicity and DMSO

Coco de Koning, Manon Beekhuijzen, Marysia Tobor-Kapłon, Selinda de Vries-Buitenweg, Dick Schoutsen, Nico Leeijen, Beppy van de Waart, Harry Emmen

Birth Defects Res B Dev Reprod Toxicol. 2015 Dec;104(6):253-72.

PMID: 26663754

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP56392177-A	(±)-Metoprolol (+)-tartrate salt		56392-17-7	Price

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