{"title":"Guide to the Larval Zebrafish-Aspergillus Infection Model.","authors":"Taylor J Schoen, Anna Huttenlocher, Nancy P Keller","doi":"10.1002/cpz1.317","DOIUrl":null,"url":null,"abstract":"<p><p>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.</p>","PeriodicalId":11174,"journal":{"name":"Current Protocols","volume":"1 12","pages":"e317"},"PeriodicalIF":0.0000,"publicationDate":"2021-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8667203/pdf/nihms-1750951.pdf","citationCount":"3","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1002/cpz1.317","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 3
Abstract
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.
斑马鱼幼虫-曲霉菌感染模型指南。
斑马鱼幼虫是一种越来越受欢迎的研究曲霉病的宿主模型。斑马鱼幼虫的视觉可及性、遗传资源、体积小和易于处理使其适合于更高通量的真菌毒力和宿主抗性机制的研究。本文提出了制备烟曲霉孢子疫苗并应用显微注射感染斑马鱼幼体后脑室所需的工艺方案。此外，我们还包括分析寄主存活、固定幼虫进行实时成像的方案，以及对图像分析的建议。©2021 Wiley期刊有限责任公司支持方案1:制备曲霉孢子支持方案2:脱毛斑马鱼胚胎支持方案3:用1-苯基2-硫脲(PTU)产生透明幼虫基本方案1:用曲霉孢子对斑马鱼幼虫进行后脑显微注射基本方案2:生存分析基本方案3:感染幼虫的多日成像备用方案:将幼虫包埋在低溶点琼脂糖中。
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