PUBLICATION

Expression profiling of the RPE in zebrafish smarca4 mutant revealed altered signals that potentially affect RPE and retinal differentiation

Authors
Zhang, L., Ma, P., Collery, R., Trowbridge, S., Zhang, M., Zhong, W., and Leung, Y.F.
ID
ZDB-PUB-140318-13
Date
2014
Source
Molecular Vision   20: 56-72 (Journal)
Registered Authors
Collery, Ross, Leung, Yuk Fai, Zhang, Liyun
Keywords
none
Datasets
GEO:GSE50241
MeSH Terms
  • Adaptor Proteins, Signal Transducing/genetics*
  • Animals
  • Cell Differentiation/genetics
  • Gene Expression Profiling*
  • Gene Expression Regulation, Developmental*
  • Melanosomes/metabolism
  • Mutation/genetics*
  • Oligonucleotide Array Sequence Analysis
  • Retinal Pigment Epithelium/embryology
  • Retinal Pigment Epithelium/metabolism
  • Retinal Pigment Epithelium/pathology*
  • Retinal Pigment Epithelium/ultrastructure
  • Signal Transduction/genetics
  • Zebrafish/embryology*
  • Zebrafish/genetics*
  • Zebrafish Proteins/genetics*
PubMed
24426776
Abstract

Purpose

The purpose of this study was to develop a framework for analyzing retinal pigment epithelium (RPE) expression profiles from zebrafish eye mutants.

Methods

The fish model we used was SWI/SNF-related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 4 (smarca4), a retinal dystrophic mutant with a previously described retinal phenotype and expression profiles. Histological and Affymetrix GeneChip analyses were conducted to characterize the RPE defects and underlying differential expression, respectively.

Results

Histological analysis revealed that smarca4 RPE was formed, but its differentiation was abnormal. In particular, ultrastructural analysis of smarca4 RPE by transmission electron microscopy demonstrated several defects in melanogenesis. The nature of these defects also suggests that the cytoskeletal dynamics, which are tightly linked with melanogenesis, were impaired in smarca4 RPE. To compare the expression profile of normal wild-type (WT) and smarca4 RPE, the gene expression profiles of microdissected retinas and RPE-attached retinas were measured with Affymetrix GeneChip analysis. The RPE expression values were then estimated from these samples by subtracting the retinal expression values from the expression values of the RPE-attached retinas. A factorial analysis was conducted using the expression values of the RPE, retinal, and whole-embryo samples. Specific rules (contrasts) were built using the coefficients of the resulting fitted models to select for three groups of genes: 1) smarca4-regulated RPE genes, 2) smarca4-regulated retinal genes, and 3) smarca4-regulated RPE genes that are not differentially expressed in the retina. Interestingly, the third group consists of 39 genes that are highly related to cytoskeletal dynamics, melanogenesis, and paracrine and intracellular signal transduction.

Conclusions

Our analytical framework provides an experimental approach to identify differentially-regulated genes in the retina and the RPE of zebrafish mutants in which both of these tissues are affected by the underlying mutation. Specifically, we have used the method to identify a group of 39 genes that can potentially explain the melanogenesis defect in the smarca4 RPE. In addition, several genes in this group are secreted signaling molecules. Thus, this observation further implicates that the smarca4 RPE might play a role in the retinal dystrophic phenotype in smarca4.

Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping