Evaluation of the pathogenesis and treatment of Mycobacterium marinum infection in zebrafish.
Evaluation of the pathogenesis and treatment of Mycobacterium marinum infection in zebrafish.
复制标题
DOI：
10.1038/nprot.2013.068
复制
发表时间：
2013-06
期刊：
影响因子：
14.8
通讯作者：
中科院分区：
文献类型：
--
作者：
感染海分枝杆菌的斑马鱼用于研究结核病发病机制以及抗结核药物的发现。斑马鱼幼虫体积小，加上其光学透明性，可以快速分析细菌负荷和宿主存活情况，以响应分枝杆菌和宿主的遗传和药理学操作。自动荧光显微镜和自动板荧光测定法 (APF) 与简便的饲养相结合，有助于对单个受感染的鱼进行大规模、重复的分析。两种方法都可以对化学库进行体内筛选，每条鱼只需要 0.1 μmol 的药物即可评估功效；它们还可以对结核病发病机制的各个阶段进行更详细的评估。在这里，我们描述了一个跨越 22 天的 16 小时方案，其中斑马鱼幼虫通过两个主要注射部位（后脑室和尾静脉）感染；接下来是发病机制和抗菌功效的高通量评估。
Mycobacterium marinum infected zebrafish are used to study tuberculosis pathogenesis, as well as for antitubercular drug discovery. The small size of zebrafish larvae coupled with their optical transparency allows for rapid analysis of bacterial burdens and host survival in response to genetic and pharmacological manipulations of both mycobacteria and host. Automated fluorescence microscopy and automated plate fluorimetry (APF) are coupled with facile husbandry to facilitate large-scale, repeated analysis of individual infected fish. Both methods allow for in vivo screening of chemical libraries, requiring only 0.1 μmol of drug per fish to assess efficacy; they also permit a more detailed evaluation of the individual stages of tuberculosis pathogenesis. Here we describe a 16-h protocol spanning 22 d, in which zebrafish larvae are infected via the two primary injection sites, the hindbrain ventricle and caudal vein; this is followed by the high-throughput evaluation of pathogenesis and antimicrobial efficacy.