Evaluation of 14 organic solvents and carriers for screening applications in zebrafish embryos and larvae.
Evaluation of 14 organic solvents and carriers for screening applications in zebrafish embryos and larvae.
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评估14个有机溶剂和载体，用于筛查斑马鱼胚胎和幼虫。
DOI：
10.1371/journal.pone.0043850
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Crawford AD
中科院分区：
文献类型：
--
作者：
Maes J;Verlooy L;Buenafe OE;de Witte PA;Esguerra CV;Crawford AD
斑马鱼作为化学遗传学、药物发现和毒理学的体内模型系统，以及最近还用于天然产品发现的模型系统，正迅速受到欢迎。在斑马鱼生物测定中，涉及小分子或天然产品提取物的药理学评价的实验需要将这些化合物有效地输送到胚胎和幼虫。虽然大多数要筛选的样品首先溶解在二甲基亚砜(DMSO)中，然后在胚胎培养液中稀释，但这种方法通常不足以防止样品立即或最终沉淀。某些化合物和提取物在DMSO中也不是高度可溶的。在这种情况下，使用载体和/或其他溶剂可提供实现所需样品浓度的替代手段。为此，我们测定了几种常用溶剂和载体在斑马鱼胚胎和仔鱼不同发育阶段的最大耐受浓度(MTC)。本研究评估的溶剂包括丙酮、乙腈、丁酮、二甲基甲酰胺、二甲基亚砜、乙醇、甘油、异丙醇、甲醇、聚乙二醇(PEG400)、丙二醇和醇缩醛，载体包括白蛋白(BSA)和环糊精(2-羟丙基-β-环糊精，或HPBCD)。这项研究的结果是确定了聚乙二醇(PEG400)、丙二醇和甲醇作为在一系列发育阶段中相对耐受的溶剂。此外，我们的结果表明，胚胎对丙酮有很好的耐受性，但幼虫对丙酮没有耐受性，1%环糊精(HPBCD)对胚胎和幼虫都有很好的耐受性，这表明该载体在斑马鱼化合物筛选中的应用。然而，鉴于表面上安全的浓度和明显有毒的浓度之间的差异相对较小(2-3倍)，应该进行进一步的研究--例如组学分析--以确定哪些细胞过程和信号通路受到用于斑马鱼小分子筛选的任何溶剂和载体的影响。
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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影响因子：
4
作者：
Bowman TV;Zon LI
通讯作者：
Zon LI
影响因子：
2.7
作者：
Blader, P;Strähle, U
通讯作者：
Strähle, U
影响因子：
3.7
作者：
Crawford AD;Liekens S;Kamuhabwa AR;Maes J;Munck S;Busson R;Rozenski J;Esguerra CV;de Witte PA
通讯作者：
de Witte PA
影响因子：
8.8
作者：
Hallare, AV;Köhler, HR;Triebskorn, R
通讯作者：
Triebskorn, R
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Feng, B;Schwarz, H;Jesuthasan, S
通讯作者：
Jesuthasan, S