Characterization of Sema6A reverse signaling in zebrafish eye development

Conference Year

January 2021

Abstract

Semaphorin6A (Sema6A) and PlexinA2 (PlxnA2) are a receptor/ligand pair involved in cell migration and axon guidance during retinal development. We previously demonstrated Sema6A and PlxnA2 to be 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 (deltaC-Sema6A) incapable of reverse signaling. The results suggest that Sema6A reverse signaling plays an important role in the maintenance of retinal integrity. Using biochemical approaches and mass spectrometry, we have identified several Sema6A binding partners as well as several phosphorylation sites on the Sema6A intracellular domain. The data suggest mechanisms whereby Sema6A reverse signaling contributes to retinal development.

Primary Faculty Mentor Name

Bryan Ballif

Secondary Mentor Name

Alicia Ebert

Faculty/Staff Collaborators

Alicia Ebert (co-PI) and Bryan Ballif (co-PI)

Status

Graduate

Student College

College of Arts and Sciences

Program/Major

Biology

Primary Research Category

Biological Sciences

Abstract only.

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Characterization of Sema6A reverse signaling in zebrafish eye development

Semaphorin6A (Sema6A) and PlexinA2 (PlxnA2) are a receptor/ligand pair involved in cell migration and axon guidance during retinal development. We previously demonstrated Sema6A and PlxnA2 to be 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 (deltaC-Sema6A) incapable of reverse signaling. The results suggest that Sema6A reverse signaling plays an important role in the maintenance of retinal integrity. Using biochemical approaches and mass spectrometry, we have identified several Sema6A binding partners as well as several phosphorylation sites on the Sema6A intracellular domain. The data suggest mechanisms whereby Sema6A reverse signaling contributes to retinal development.