PUBLICATION

Loss of foxc1 in zebrafish reduces optic nerve size and cell number in the ganglion cell layer

Authors
Umali, J., Hawkey-Noble, A., French, C.R.
ID
ZDB-PUB-190127-18
Date
2019
Source
Vision Research   156: 66-72 (Journal)
Registered Authors
Keywords
CRISPR, Glaucoma, Retinal Ganglion Cells, Zebrafish, atoh7, foxc1, pou4f2 FGF19
MeSH Terms
  • Animals
  • Axons/pathology
  • Cell Count
  • Cell Death
  • Cell Differentiation
  • DNA-Binding Proteins/genetics
  • Embryo, Nonmammalian
  • Forkhead Transcription Factors/genetics*
  • Gene Expression Regulation, Developmental/physiology*
  • Gene Silencing/drug effects
  • Glaucoma/embryology
  • Glaucoma/genetics
  • In Situ Hybridization
  • Morpholinos/pharmacology
  • Mutation*
  • Optic Nerve/embryology*
  • Polymerase Chain Reaction
  • Retinal Ganglion Cells/pathology*
  • Transfection
  • Zebrafish/embryology*
  • Zebrafish Proteins/genetics*
PubMed
30684501 Full text @ Vision Res.
Abstract
Mutation of FOXC1 causes Axenfeld-Rieger Syndrome (ARS) with early onset or congenital glaucoma. We assessed retinal ganglion cell (RGC) number in zebrafish due to CRISPR-mediated mutation and antisense inhibition of two-forkhead box transcription factors, foxc1a and foxc1b. These genes represent duplicated homologues of human FOXC1. Using a CRISPR induced null mutation in foxc1b, in combination with antisense inhibition of foxc1a, we demonstrate reduced cell number in the retinal ganglion cell layer of developing zebrafish eyes. As early as 5 days post fertilization (dpf), fewer RGCs are found in foxc1b homozygous mutants injected with foxc1a morpholinos, and a thinner optic nerve results. Our data illustrates that foxc1 is required for the expression of atonal homolog 7 (atoh7), a gene that is necessary for RGC differentiation. As markers of differentiated RGCs (pou4f2) are downregulated in foxc1b-/- mutants injected with foxc1a morpholinos and no cell death is observed, our results are consistent with defects in the differentiation of RGCs leading to reduced cell number, as opposed to increased cell death of RGCs or off targets effects of morpholino injection. Our zebrafish model demonstrates that aberrant regulation of RGC number could act in concert with other known glaucoma risk factors to influence the development of congenital and early onset glaucoma due to FOXC1 mutation.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping