斑马鱼(Danio rerio)胚胎的发育阶段和使用 Foldscope 显微镜进行的毒理学研究。

Developmental stages of zebrafish (Danio rerio) embryos and toxicological studies using foldscope microscope.

机构信息

Regeneration and Stem Cell Biology Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science and Technology, Chennai, Tamilnadu, India.

出版信息

Cell Biol Int. 2020 Oct;44(10):1968-1980. doi: 10.1002/cbin.11412. Epub 2020 Jul 1.

Abstract

Zebrafish (Danio rerio), is a well-established vertebrate animal model widely used in developmental biology and toxicological research. In the present study, foldscope is used as an innovative tool to study the developmental stages and toxicological analysis of the zebrafish embryos. Briefly, the developmental stages, such as zygote, cleavage, blastula, gastrula, segmentation, and pharyngula formation are observed and documented using simple foldscope. Toxicological parameters upon exposure to different concentration of ethanol extract of Curcuma longa and its lead compound, ar-turmerone along with rhodamine B (bio-coupler) on zebrafish embryos are analyzed upto 72 hr using foldscopes in live condition. The lethal endpoints, such as coagulation, lack of somite formation, non-detachment of tail, and lack of heartbeat are clearly monitored and documented using foldscope. Bio-evaluation of test compounds with the aid of foldscope confirms that the toxicity is directly proportional to the concentration. Our results conclude that, ethanol extract of C. longa, ar-turmerone and rhodamine B exposed embryos remains healthy up to 96, 48, and 24 µg concentrations, respectively. Embryos exposed to higher concentrations become coagulated, however normal physiological active movement of tail lashing and heartbeat are evident in lower concentration exposed embryos. Except coagulation, no other abnormalities are observed and interestingly, the hatching ability is not delayed, when compared with the control embryos. It is confirmed that the test compounds are not highly toxic to zebrafish embryos. Hence it can be used for further analysis, especially for studying the neural-regeneration and its neuronal development in zebrafish embryos.

摘要

斑马鱼(Danio rerio)是一种成熟的脊椎动物模型,广泛应用于发育生物学和毒理学研究。在本研究中,foldscope 被用作一种创新工具来研究斑马鱼胚胎的发育阶段和毒理学分析。简而言之,使用简单的 foldscope 观察和记录发育阶段,如受精卵、卵裂、囊胚、原肠胚、分节和咽形成。暴露于不同浓度的姜黄乙醇提取物及其主要化合物莪术烯以及罗丹明 B(生物偶联剂)对斑马鱼胚胎的毒理学参数在活体内用 foldscope 分析至 72 小时。使用 foldscope 清楚地监测和记录致死终点,如凝固、体节形成缺失、尾部未分离和无心跳。在 foldscope 的辅助下对测试化合物进行生物评估,确认毒性与浓度成正比。我们的结果表明,姜黄乙醇提取物、莪术烯和罗丹明 B 暴露的胚胎在 96、48 和 24µg 浓度下仍然保持健康。暴露于较高浓度的胚胎会凝固,但在较低浓度暴露的胚胎中,尾部鞭打的正常生理活动和心跳是明显的。除凝固外,未观察到其他异常,有趣的是,与对照胚胎相比,孵化能力没有延迟。可以确认测试化合物对斑马鱼胚胎没有高度毒性。因此,它可以用于进一步的分析,特别是研究斑马鱼胚胎中的神经再生及其神经元发育。

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