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

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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.