《斑马鱼幼虫 - 曲霉菌感染模型指南》。
Guide to the Larval Zebrafish-Aspergillus Infection Model.
机构信息
Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, Wisconsin.
Comparative Biomedical Sciences Graduate Program, University of Wisconsin-Madison, Madison, Wisconsin.
出版信息
Curr Protoc. 2021 Dec;1(12):e317. doi: 10.1002/cpz1.317.
The larval zebrafish is an increasingly popular host model for the study of Aspergillosis. The visual accessibility, genetic resources, small size, and ease of handling make zebrafish larvae compatible with higher-throughput investigation of fungal virulence and host resistance mechanisms. This article provides the protocols needed to prepare Aspergillus fumigatus spore inocula and use microinjection to infect the hindbrain ventricle of zebrafish larvae. Furthermore, we include protocols for analyzing host survival, immobilizing larvae for live imaging, and suggestions for image analysis. © 2021 Wiley Periodicals LLC. Support Protocol 1: Preparing Aspergillus spores Support Protocol 2: Dechorionating zebrafish embryos Support Protocol 3: Generating transparent larvae with 1-phenyl 2-thiourea (PTU) Basic Protocol 1: Hindbrain microinjection of zebrafish larvae with Aspergillus spores Basic Protocol 2: Survival analysis Basic Protocol 3: Multi-day imaging of infected larvae Alternate Protocol: Embedding larvae in low-melting-point agarose.
幼虫斑马鱼是研究曲霉病的一种越来越受欢迎的宿主模型。其视觉可及性、遗传资源、体型小和易于操作，使斑马鱼幼虫适合于进行真菌毒力和宿主抗性机制的高通量研究。本文提供了制备烟曲霉孢子接种物和使用显微注射感染斑马鱼幼虫后脑室所需的方案。此外，我们还包括了用于分析宿主存活、固定幼虫进行活体成像以及图像分析建议的方案。© 2021 Wiley Periodicals LLC. 支持方案 1：制备曲霉孢子 支持方案 2：去膜斑马鱼胚胎 支持方案 3：用 1-苯基 2-硫脲（PTU）生成透明幼虫 基本方案 1：用曲霉孢子对斑马鱼幼虫进行后脑微注射 基本方案 2：存活分析 基本方案 3：感染幼虫的多天成像 备选方案：将幼虫嵌入低熔点琼脂糖中。
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