Large-scale analysis of acute ethanol exposure in zebrafish development: a critical time window and resilience.
Large-scale analysis of acute ethanol exposure in zebrafish development: a critical time window and resilience.
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DOI：
10.1371/journal.pone.0020037
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Richardson MK
中科院分区：
文献类型：
--
作者：
Ali S;Champagne DL;Alia A;Richardson MK
在人类中，怀孕期间暴露于乙醇会导致一系列发育缺陷（胎儿酒精综合征或FAS）。个体的表型表达各不相同。斑马鱼胚胎乙醇暴露后发展FAS样功能。在这项研究中，我们问是否可以在斑马鱼中确定乙醇的特定阶段的影响，如果是这样，它们是否允许精确定位敏感的发育机制。因此，我们进行了第一次大规模（>1500个胚胎）分析急性，阶段特异性药物对斑马鱼发育的影响，并获得了大量的读数。斑马鱼胚胎在96孔板中培养。范围探索表明，10%乙醇持续1 h适用于急性暴露方案。高分辨率魔角旋转质子磁共振波谱显示，这在绒毛膜内的胚胎中产生了浓度为0.86%的乙醇的瞬时脉冲。对受精后5天的存活者进行了分析。表型范围从正常（弹性）到严重畸形。早期乙醇暴露导致高死亡率（≥88%）。在接触的后期阶段，死亡率下降，并出现畸形。咽弓发育不全和行为障碍在接触α-6和α-16后最为常见。相比之下，小眼球和生长迟缓是阶段无关的。我们的研究结果表明，一些乙醇的影响是强烈的阶段依赖性。表型模拟FAS的关键方面，包括颅面异常，小眼，生长迟缓和行为障碍。我们还确定了乙醇敏感性的关键时间窗口（10 -6和10 -16）。最后，我们鉴定了一个广泛的表型谱，让人想起人类FAS，并可能提供一个有用的模型，研究疾病的弹性。
In humans, ethanol exposure during pregnancy causes a spectrum of developmental defects (fetal alcohol syndrome or FAS). Individuals vary in phenotypic expression. Zebrafish embryos develop FAS-like features after ethanol exposure. In this study, we ask whether stage-specific effects of ethanol can be identified in the zebrafish, and if so, whether they allow the pinpointing of sensitive developmental mechanisms. We have therefore conducted the first large-scale (>1500 embryos) analysis of acute, stage-specific drug effects on zebrafish development, with a large panel of readouts. Zebrafish embryos were raised in 96-well plates. Range-finding indicated that 10% ethanol for 1 h was suitable for an acute exposure regime. High-resolution magic-angle spinning proton magnetic resonance spectroscopy showed that this produced a transient pulse of 0.86% concentration of ethanol in the embryo within the chorion. Survivors at 5 days postfertilisation were analysed. Phenotypes ranged from normal (resilient) to severely malformed. Ethanol exposure at early stages caused high mortality (≥88%). At later stages of exposure, mortality declined and malformations developed. Pharyngeal arch hypoplasia and behavioral impairment were most common after prim-6 and prim-16 exposure. By contrast, microphthalmia and growth retardation were stage-independent. Our findings show that some ethanol effects are strongly stage-dependent. The phenotypes mimic key aspects of FAS including craniofacial abnormality, microphthalmia, growth retardation and behavioral impairment. We also identify a critical time window (prim-6 and prim-16) for ethanol sensitivity. Finally, our identification of a wide phenotypic spectrum is reminiscent of human FAS, and may provide a useful model for studying disease resilience.