A method for the isolation of miRNAs from tick ex vivo salivary gland cultures and extracellular vesicles

Authors: LEAL, BRENDA - Texas A&M University; HARVEY, CRISTINA - Texas A&M University; Thomas, Donald; OLIVA-CHAVEZ, ADELA - Texas A&M University

Submitted to: The Journal of Visualized Experiments (JoVE)
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/24/2022
Publication Date: 4/26/2022
Citation: Leal, B., Harvey, C., Thomas, D.B., Oliva-Chavez, A. 2022. A method for the isolation of miRNAs from tick ex vivo salivary gland cultures and extracellular vesicles. The Journal of Visualized Experiments (JoVE). https://doi.org/10.3791/63618.
DOI: https://doi.org/10.3791/63618

Interpretive Summary: Ticks can spread disease through their bites. The saliva of the ticks contain pharmaceutical properties that can diminish host immune responses and enhance pathogen transmission. One group of such compounds are small RNAs (miRNAs). miRNAs are short non-coding sequences that can regulate host gene expression at the tick-host interface and within the organs of the tick. These small RNAs can be transported in the tick saliva via tiny blebs called extracellular vesicles (EVs), which pass materials between one cell and another. Vesicles containing small RNAs have been described in the saliva of ticks. However, little is known about the specific actions by these small RNAs in tick salivary vesicles and glands. Furthermore, the study of vesicles and small RNAs in tick saliva requires tedious procedures for the collection of tick saliva. The focus of this project is to develop and validate a method for isolating purified extracellular vesicles and small RNAs from tick organs. Herein, we describe the materials and methodology needed to extract small RNAs from extracellular vesicles and tick salivary glands.

Technical Abstract: Ticks are important ectoparasites that can vector multiple pathogens. The salivary glands of the ticks are essential for feeding as it contains many effectors with pharmaceutical properties that can diminish host immune responses and enhance pathogen transmission. One group of such effectors are microRNAs (miRNAs). miRNAs are short non-coding sequences that can regulate host gene expression at the tick-host interface and within the organs of the tick. These small RNAs can be transported in the tick saliva via extracellular vesicles (EVs), which serve in inter- and intra-cellular communication. Vesicles containing miRNAs have been described in the saliva of ticks. However, little is known about the roles and profiles of the miRNAs in tick salivary vesicles and glands. Furthermore, the study of vesicles and miRNAs in tick saliva requires tedious procedures for the collection of tick saliva. The focus of this project is to develop and validate a method for isolating purified extracellular vesicles and miRNAs from ex vivo organ cultures. Herein, we describe the materials and methodology needed to extract miRNAs from extracellular vesicles and tick salivary glands.