Researcher(s)
- Krisha Parekh, Biological Sciences, University of Delaware
Faculty Mentor(s)
- Jessica Tanis, Biological Sciences, University of Delaware
Abstract
Extracellular vesicles (EVs) are lipid-bound vesicles that can transport various biological macromolecules and act in communication between cells. In Caenorhabditis elegans, EVs are shed from the primary cilia, which are organelles that play a role in signal transduction and transmission. In fact, two specific ciliary EV cargoes are released, one that contains the TRP polycystin channel PKD-2 and another that contains the ion channel CLHM-1. To visualize the EVs that have been shed, CLHM has been tagged with tdTomato, and PKD has been tagged with green fluorescent protein (GFP). Prior research indicates that these EV subpopulations are differentially shed, as PKD-2 EVs are released from the ciliary distal tip, while CLHM-1 EVs are released from the ciliary base. I am investigating the effects of the loss of stam-1 on the shedding of CLHM-1 EVs, PKD-2 EVs, or both subpopulations. stam-1 is a protein that is part of the endosomal recycling complex required for transport (ESCRT) machinery, which has previously been found to play a role in EV biogenesis. The protein is also known for its involvement in the downregulation of PKD-2. Genetic crosses with C. elegans first had to be formed to produce nematodes that are homozygous for the stam-1 mutation and the transgenes that express the EV cargoes tagged with fluorescent proteins. Since the desired strain has been isolated, the worms are currently being imaged to determine the ciliary localization of CLHM-1::tdT and PKD-2::GFP when the stam-1 gene is present and absent, as well as quantify the amount of EVs that are released by stam-1 mutants and wildtype. Through this research, there may be a greater understanding of the mechanisms that regulate shedding of EV subpopulations.