Differential Gene Expression in Danio rerio during Optic Nerve Regeneration
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Spinal cord injuries and neurodegenerative diseases in mammals result in a loss of function due to the failure of neurons in the central nervous system (CNS) to survive and regenerate their axons. Unlike mammals, fish and amphibians possess the ability to regenerate their CNS following damage. To gain a better understanding of the factors necessary for successful CNS regeneration, I conducted a temporal analysis of the changes in gene expression in the retina caused by optic nerve injury to identify genes specifically involved in regeneration. Dual color oligonucleotide microarrays were used to compare total RNA harvested from retinas of sham-operated and optic nerve-injured fish at 3, 24 and 168 hours following surgery. Statistical analyses identified 722 genes differentially expressed by at least 1.5-fold at one or more time points, and 142 genes with at least a 2.0-fold difference. Based on microarray fold differences and gene ontology analysis, six genes were selected for further analysis using qRT-PCR. The results of qRT-PCR identified noggin 2, activating transcription factor 3, and beta-tubulin 5 as genes that showed significantly increased expression in the injured fish as compared to sham; therefore, these genes’ products may play an important role in optic nerve regeneration in zebrafish. These results support the hypothesis that an analysis of gene expression between optic nerve injured and sham-operated fish will reveal genes specifically involved in regeneration.