Characterization of forward and reverse Sema6A signaling in Zebrafish eye development
Conference Year
January 2019
Abstract
Semaphorin6A (Sema6A) and PlexinA2 (PlxnA2) are a receptor/ligand pair involved in cell migration and axon guidance of neurons and blood vessels, among other cell types. We previously demonstrated Sema6a and PlxnA2 as an essential signaling pair for zebrafish eye development, as knockdown of either leads to cell adhesion and proliferation defects. It is known that Sema6A can act both as a ligand in canonical downstream signaling through the PlxnA2 receptor, and as a receptor in reverse signaling. In order to determine the importance of the reverse Sema6A-PlxnA2 signaling in zebrafish eye development, we knocked down Sema6A with an antisense oligonucleotide and rescued with full length human Sema6A (FL-Sema6A) or a truncated Sema6A without the intracellular domain (DC-Sema6A) incapable of reverse signaling. Using biochemistry techniques, we have shown that FL-Sema6A and DC-Sema6A are localized similarly in the cell but are processed unevenly leading to a soluble protein that is released into the media. We are currently investigating the ability of DC-Sema6A and the soluble Sema6A to rescue the eye phenotypes observed with loss of endogenous Sema6A. These experiments will lend insight into how reverse signaling and the soluble Sema6A are involved in development of the zebrafish eye.
Primary Faculty Mentor Name
Alicia Ebert
Secondary Mentor Name
Bryan Ballif
Graduate Student Mentors
Riley St. Clair
Faculty/Staff Collaborators
Riley M. St. Clair, Abagael M. Lasseigne
Status
Undergraduate
Student College
College of Arts and Sciences
Program/Major
Neuroscience
Primary Research Category
Biological Sciences
Characterization of forward and reverse Sema6A signaling in Zebrafish eye development
Semaphorin6A (Sema6A) and PlexinA2 (PlxnA2) are a receptor/ligand pair involved in cell migration and axon guidance of neurons and blood vessels, among other cell types. We previously demonstrated Sema6a and PlxnA2 as an essential signaling pair for zebrafish eye development, as knockdown of either leads to cell adhesion and proliferation defects. It is known that Sema6A can act both as a ligand in canonical downstream signaling through the PlxnA2 receptor, and as a receptor in reverse signaling. In order to determine the importance of the reverse Sema6A-PlxnA2 signaling in zebrafish eye development, we knocked down Sema6A with an antisense oligonucleotide and rescued with full length human Sema6A (FL-Sema6A) or a truncated Sema6A without the intracellular domain (DC-Sema6A) incapable of reverse signaling. Using biochemistry techniques, we have shown that FL-Sema6A and DC-Sema6A are localized similarly in the cell but are processed unevenly leading to a soluble protein that is released into the media. We are currently investigating the ability of DC-Sema6A and the soluble Sema6A to rescue the eye phenotypes observed with loss of endogenous Sema6A. These experiments will lend insight into how reverse signaling and the soluble Sema6A are involved in development of the zebrafish eye.