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A Novel Light Damage Paradigm for Use in Retinal Regeneration Studies in Adult Zebrafish
Published on: October 24, 2013
Microarray analysis of XOPS-mCFP zebrafish retina identifies genes associated with rod photoreceptor degeneration and
Ann C Morris1, Marie A Forbes-Osborne, Lakshmi S Pillai
1Department of Biology, University of Kentucky, Lexington, KY 40506, USA. ann.morris@uky.edu
Purpose:
XOPS-mCFP transgenic zebrafish experience a continual cycle of rod photoreceptor development and degeneration throughout life, making them a useful model for investigating the molecular determinants of rod photoreceptor regeneration. The purpose of this study was to compare the gene expression profiles of wild-type and XOPS-mCFP retinas and identify genes that may contribute to the regeneration of the rods.
Methods:
Adult wild-type and XOPS-mCFP retinal mRNA was subjected to microarray analysis. Pathway analysis was used to identify biologically relevant processes that were significantly represented in the dataset. Expression changes were verified by RT-PCR. Selected genes were further examined during retinal development and in adult retinas by in situ hybridization and immunohistochemistry and in a transgenic fluorescent reporter line.
Results:
More than 600 genes displayed significant expression changes in XOPS-mCFP retinas compared with expression in wild-type controls. Many of the downregulated genes were associated with phototransduction, whereas upregulated genes were associated with several biological functions, including cell cycle, DNA replication and repair, and cell development and death. RT-PCR analysis of a subset of these genes confirmed the microarray
Results:
Three transcription factors (sox11b, insm1a, and c-myb), displaying increased expression in XOPS-mCFP retinas, were also expressed throughout retinal development and in the persistently neurogenic ciliary marginal zone.
Conclusions:
This study identified numerous gene expression changes in response to rod degeneration in zebrafish and further suggests a role for the transcriptional regulators sox11b, insm1a, and c-myb in both retinal development and rod photoreceptor regeneration.
Insights
Zebrafish with continuously regenerating rods reveal over 600 gene expression changes. Key transcription factors like sox11b, insm1a, and c-myb are implicated in rod development and regeneration.
Area of Science:
- * Retinal biology and molecular genetics.
- * Photoreceptor development and regeneration.
Background:
- * XOPS-mCFP transgenic zebrafish exhibit ongoing rod photoreceptor development and degeneration.
- * This model is valuable for studying the molecular mechanisms underlying rod regeneration.
Purpose of the Study:
- * To compare gene expression profiles between wild-type and XOPS-mCFP zebrafish retinas.
- * To identify genes potentially involved in rod photoreceptor regeneration.
Main Methods:
- * Microarray analysis of adult wild-type and XOPS-mCFP retinal mRNA.
- * Pathway analysis to identify significant biological processes.
- * Validation of expression changes using RT-PCR, in situ hybridization, and immunohistochemistry.
Main Results:
- * Over 600 genes showed significant expression differences in XOPS-mCFP retinas.
- * Downregulated genes were linked to phototransduction; upregulated genes to cell cycle, DNA repair, and cell death/development.
- * Increased expression of transcription factors sox11b, insm1a, and c-myb was observed.
Conclusions:
- * Numerous gene expression alterations occur in response to rod degeneration in zebrafish.
- * The transcription factors sox11b, insm1a, and c-myb are suggested to play roles in retinal development and rod regeneration.
