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Evaluation of 14 Organic Solvents and Carriers for Screening Applications in Zebrafish Embryos and Larvae

Zebrafish are rapidly growing in popularity as an in vivo model system for chemical genetics, drug discovery, and toxicology, and more recently also for natural product discovery. Experiments involving the pharmacological evaluation of small molecules or natural product extracts in zebrafish bioassa...

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Autores principales: Maes, Jan, Verlooy, Lien, Buenafe, Olivia E., de Witte, Peter A. M., Esguerra, Camila V., Crawford, Alexander D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3474771/
https://www.ncbi.nlm.nih.gov/pubmed/23082109
http://dx.doi.org/10.1371/journal.pone.0043850
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author Maes, Jan
Verlooy, Lien
Buenafe, Olivia E.
de Witte, Peter A. M.
Esguerra, Camila V.
Crawford, Alexander D.
author_facet Maes, Jan
Verlooy, Lien
Buenafe, Olivia E.
de Witte, Peter A. M.
Esguerra, Camila V.
Crawford, Alexander D.
author_sort Maes, Jan
collection PubMed
description Zebrafish are rapidly growing in popularity as an in vivo model system for chemical genetics, drug discovery, and toxicology, and more recently also for natural product discovery. Experiments involving the pharmacological evaluation of small molecules or natural product extracts in zebrafish bioassays require the effective delivery of these compounds to embryos and larvae. While most samples to be screened are first solubilized in dimethyl sulfoxide (DMSO), which is then diluted in the embryo medium, often this method is not sufficient to prevent the immediate or eventual precipitation of the sample. Certain compounds and extracts are also not highly soluble in DMSO. In such instances the use of carriers and/or other solvents might offer an alternative means to achieve the required sample concentration. Towards this end, we determined the maximum tolerated concentration (MTC) of several commonly used solvents and carriers in zebrafish embryos and larvae at various developmental stages. Solvents evaluated for this study included acetone, acetonitrile, butanone, dimethyl formamide, DMSO, ethanol, glycerol, isopropanol, methanol, polyethylene glycol (PEG-400), propylene glycol, and solketal, and carriers included albumin (BSA) and cyclodextrin (2-hydroxypropyl-beta-cyclodextrin, or HPBCD). This study resulted in the identification of polyethylene glycol (PEG400), propylene glycol, and methanol as solvents that were relatively well-tolerated over a range of developmental stages. In addition, our results showed that acetone was well-tolerated by embryos but not by larvae, and 1% cyclodextrin (HPBCD) was well-tolerated by both embryos and larvae, indicating the utility of this carrier for compound screening in zebrafish. However, given the relatively small differences (2–3 fold) between concentrations that are apparently safe and those that are clearly toxic, further studies – e.g. omics analyses –should be carried out to determine which cellular processes and signalling pathways are affected by any solvents and carriers that are used for small-molecule screens in zebrafish.
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spelling pubmed-34747712012-10-18 Evaluation of 14 Organic Solvents and Carriers for Screening Applications in Zebrafish Embryos and Larvae Maes, Jan Verlooy, Lien Buenafe, Olivia E. de Witte, Peter A. M. Esguerra, Camila V. Crawford, Alexander D. PLoS One Research Article Zebrafish are rapidly growing in popularity as an in vivo model system for chemical genetics, drug discovery, and toxicology, and more recently also for natural product discovery. Experiments involving the pharmacological evaluation of small molecules or natural product extracts in zebrafish bioassays require the effective delivery of these compounds to embryos and larvae. While most samples to be screened are first solubilized in dimethyl sulfoxide (DMSO), which is then diluted in the embryo medium, often this method is not sufficient to prevent the immediate or eventual precipitation of the sample. Certain compounds and extracts are also not highly soluble in DMSO. In such instances the use of carriers and/or other solvents might offer an alternative means to achieve the required sample concentration. Towards this end, we determined the maximum tolerated concentration (MTC) of several commonly used solvents and carriers in zebrafish embryos and larvae at various developmental stages. Solvents evaluated for this study included acetone, acetonitrile, butanone, dimethyl formamide, DMSO, ethanol, glycerol, isopropanol, methanol, polyethylene glycol (PEG-400), propylene glycol, and solketal, and carriers included albumin (BSA) and cyclodextrin (2-hydroxypropyl-beta-cyclodextrin, or HPBCD). This study resulted in the identification of polyethylene glycol (PEG400), propylene glycol, and methanol as solvents that were relatively well-tolerated over a range of developmental stages. In addition, our results showed that acetone was well-tolerated by embryos but not by larvae, and 1% cyclodextrin (HPBCD) was well-tolerated by both embryos and larvae, indicating the utility of this carrier for compound screening in zebrafish. However, given the relatively small differences (2–3 fold) between concentrations that are apparently safe and those that are clearly toxic, further studies – e.g. omics analyses –should be carried out to determine which cellular processes and signalling pathways are affected by any solvents and carriers that are used for small-molecule screens in zebrafish. Public Library of Science 2012-10-17 /pmc/articles/PMC3474771/ /pubmed/23082109 http://dx.doi.org/10.1371/journal.pone.0043850 Text en © 2012 Maes et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Maes, Jan
Verlooy, Lien
Buenafe, Olivia E.
de Witte, Peter A. M.
Esguerra, Camila V.
Crawford, Alexander D.
Evaluation of 14 Organic Solvents and Carriers for Screening Applications in Zebrafish Embryos and Larvae
title Evaluation of 14 Organic Solvents and Carriers for Screening Applications in Zebrafish Embryos and Larvae
title_full Evaluation of 14 Organic Solvents and Carriers for Screening Applications in Zebrafish Embryos and Larvae
title_fullStr Evaluation of 14 Organic Solvents and Carriers for Screening Applications in Zebrafish Embryos and Larvae
title_full_unstemmed Evaluation of 14 Organic Solvents and Carriers for Screening Applications in Zebrafish Embryos and Larvae
title_short Evaluation of 14 Organic Solvents and Carriers for Screening Applications in Zebrafish Embryos and Larvae
title_sort evaluation of 14 organic solvents and carriers for screening applications in zebrafish embryos and larvae
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3474771/
https://www.ncbi.nlm.nih.gov/pubmed/23082109
http://dx.doi.org/10.1371/journal.pone.0043850
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