Ixodid Tick Dissection and Tick Ex Vivo Organ Cultures for Tick-Borne Virus Research

Abstract

Tick-borne viruses cause thousands of cases of disease worldwide every year. Specific countermeasures to many tick-borne viruses are not commercially available. Very little is known regarding tick-virus interactions and increasing this knowledge can lead to potential targets for countermeasure development. Virus infection of ex vivo organ cultures from ticks can provide an approach to identify susceptible cell types of tissue to infection. Additionally, these organ cultures can be used for functional genomic studies to pinpoint tick-specific genes involved in the virus lifecycle. Provided here are step-by-step procedures to set up basic tick organ cultures in combination with virus infection and/or functional genomic studies. These procedures can be adapted for future use to characterize other tick-borne pathogen infections as well as tick-specific biological processes. © 2020 Wiley Periodicals LLC.

Basic Protocol 1: Loading 96-well plates with gelfoam substrate

Basic Protocol 2: Step-by-step aseptic dissection of unfed female/male Ixodes scapularis ticks for multiple organs

Basic Protocol 3: Step-by-step aseptic dissection of fed female Ixodes scapularis ticks to remove salivary glands

Basic Protocol 4: Metabolic viability analyses of tick organ cultures

Basic Protocol 5: Virus infection of tick organ cultures

Basic Protocol 6: Functional RNA interference analyses using tick organ cultures