PUBLICATION
Loss of Zebrafish Mfrp Causes Nanophthalmia, Hyperopia, and Accumulation of Subretinal Macrophages
- Authors
- Collery, R.F., Volberding, P.J., Bostrom, J.R., Link, B.A., Besharse, J.C.
- ID
- ZDB-PUB-161222-1
- Date
- 2016
- Source
- Investigative ophthalmology & visual science 57: 6805-6814 (Journal)
- Registered Authors
- Besharse, Joseph C., Collery, Ross, Link, Brian, Volberding, Peter
- Keywords
- none
- MeSH Terms
-
- Animals
- DNA/genetics
- DNA Mutational Analysis
- Glycoproteins/genetics*
- Glycoproteins/metabolism
- Humans
- Hyperopia/genetics*
- Hyperopia/metabolism
- Hyperopia/pathology
- Macrophages/pathology*
- Microphthalmos/genetics*
- Microphthalmos/metabolism
- Microphthalmos/pathology
- Mutation*
- Phenotype
- Polymerase Chain Reaction
- Retinal Degeneration/genetics*
- Retinal Degeneration/metabolism
- Retinal Degeneration/pathology
- Retinal Pigment Epithelium/metabolism
- Retinal Pigment Epithelium/pathology*
- Tomography, Optical Coherence
- Zebrafish
- PubMed
- 28002843 Full text @ Invest. Ophthalmol. Vis. Sci.
Citation
Collery, R.F., Volberding, P.J., Bostrom, J.R., Link, B.A., Besharse, J.C. (2016) Loss of Zebrafish Mfrp Causes Nanophthalmia, Hyperopia, and Accumulation of Subretinal Macrophages. Investigative ophthalmology & visual science. 57:6805-6814.
Abstract
Purpose Mutations in membrane frizzled-related protein (MFRP) are associated with nanophthalmia, hyperopia, foveoschisis, irregular patches of RPE atrophy, and optic disc drusen in humans. Mouse mfrp mutants show retinal degeneration but no change in eye size or refractive state. The goal of this work was to generate zebrafish mutants to investigate the loss of Mfrp on eye size and refractive state, and to characterize other phenotypes observed.
Methods Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 methods were used to generate multiple frameshift mutations in zebrafish mfrp causing premature translational stops in Mfrp. Spectral-domain optical coherence tomography (SD-OCT) was used to measure eye metrics and refractive state, and immunohistochemistry was used to study adult eyes. Gene expression levels were measured using quantitative PCR.
Results Zebrafish Mfrp was shown to localize to apical and basal regions of RPE cells, as well as the ciliary marginal zone. Loss of Mfrp in mutant zebrafish was verified histologically. Zebrafish eyes that were mfrp mutant showed reduced axial length causing hyperopia, RPE folding, and macrophages were observed subretinally. Visual acuity was reduced in mfrp mutant animals.
Conclusions Mutation of zebrafish mfrp results in hyperopia with subretinal macrophage infiltration, phenocopying aspects of human and mouse Mfrp deficiency. These mutant zebrafish will be useful in studying the onset and progression of Mfrp-related nanophthalmia, the cues that initiate the recruitment of macrophages, and the mechanisms of Mfrp function.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping