Journal
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
Volume 45, Issue 8, Pages 2737-2746Publisher
ASSOC RESEARCH VISION OPHTHALMOLOGY INC
DOI: 10.1167/iovs.03-1047
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Funding
- NEI NIH HHS [R01EY12369, R01 EY012369, P30 EY013080-04, R01 EY012369-01A1, P30 EY013080] Funding Source: Medline
- PHS HHS [5P30 E7013080-04S1] Funding Source: Medline
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PURPOSE. The goal of this study was to define the temporal changes in gene expression after retinal injury and to relate these changes to the inflammatory and reactive response. A specific emphasis was placed on the tetraspanin family of proteins and their relationship with markers of reactive gliosis. METHODS. Retinal tears were induced in adult rats by scraping the retina with a needle. After different survival times ( 4 hours, and 1, 3, 7, and 30 days), the retinas were removed, and mRNA was isolated, prepared, and hybridized to the Affymatrix RG-U34A microarray ( Santa Clara, CA). Microarray results were confirmed by using RT-PCR and correlation to protein levels was determined. RESULTS. Of the 8750 genes analyzed, approximately 393 (4.5%) were differentially expressed. Clustering analysis revealed three major profiles: ( 1) The early response was characterized by the upregulation of transcription factors; ( 2) the delayed response included a high percentage of genes related to cell cycle and cell death; and ( 3) the late, sustained profile clustered a significant number of genes involved in retinal gliosis. The late, sustained cluster also contained the upregulated crystallin genes. The tetraspanins Cd9, Cd81, and Cd82 were also associated with the late, sustained response. CONCLUSIONS. The use of microarray technology enables definition of complex genetic changes underlying distinct phases of the cellular response to retinal injury. The early response clusters genes associate with the transcriptional regulation of the wound-healing process and cell death. Most of the genes in the late, sustained response appear to be associated with reactive gliosis.
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