PUBLICATION
Infection of Zebrafish Larvae with Aspergillus Spores for Analysis of Host-Pathogen Interactions
- Authors
- Thrikawala, S., Rosowski, E.E.
- ID
- ZDB-PUB-200603-4
- Date
- 2020
- Source
- Journal of visualized experiments : JoVE (159): (Journal)
- Registered Authors
- Rosowski, Emily E., Thrikawala, Savini
- Keywords
- none
- MeSH Terms
-
- Animals
- Aspergillosis/immunology*
- Disease Models, Animal*
- Host-Pathogen Interactions*
- Immunosuppressive Agents
- Larva/microbiology*
- Spores, Fungal/immunology
- Zebrafish/microbiology*
- PubMed
- 32478760 Full text @ J. Vis. Exp.
Citation
Thrikawala, S., Rosowski, E.E. (2020) Infection of Zebrafish Larvae with Aspergillus Spores for Analysis of Host-Pathogen Interactions. Journal of visualized experiments : JoVE. (159):.
Abstract
Invasive aspergillosis (IA) is one of the most common fungal infections among immunocompromised individuals. Despite the availability of antifungal drugs, IA can cause >50% mortality in infected immunocompromised patients. It is crucial to determine both host and pathogen factors that contribute to infection susceptibility and low survival rates in infected patients in order to develop novel therapeutics. Innate immune responses play a pivotal role in recognition and clearance of Aspergillus spores, though little is known about the exact cellular and molecular mechanisms. Reliable models are required to investigate detailed mechanistic interactions between the host and pathogen. The optical clarity and genetic tractability of zebrafish larvae make them an intriguing model to study host-pathogen interactions of multiple human bacterial and fungal infections in a live and intact host. This protocol describes a larval zebrafish Aspergillus infection model. First, Aspergillus spores are isolated and injected into the zebrafish hindbrain ventricle via microinjection. Then, chemical inhibitors such as immunosuppressive drugs are added directly to the larval water. Two methods to monitor the infection in injected larvae are described, including the 1) homogenization of larvae for colony forming unit (CFU) enumeration and 2) a repeated, daily live imaging setup. Overall, these techniques can be used to mechanistically analyze the progression of Aspergillus infection in vivo and can be applied to different host backgrounds and Aspergillus strains to interrogate host-pathogen interactions.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping