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L. A. Kim, Z. Chen, B. Thomas, D. Mock, S. R. Sadda; Gene Expression Patterns of Retinal Progenitor Cells Before and After Transplantation. Invest. Ophthalmol. Vis. Sci. 2008;49(13):4277.
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© ARVO (1962-2015); The Authors (2016-present)
To analyze the gene expression patterns of retinal progenitor cells prior to and after transplantation in S334ter-line-3 (line-3) rats, a transgenic model of retinal photoreceptor degeneration.
Retinal progenitor cells (RPC) were collected from embryonic day 19 (E19) hPAP/GFP rats. These RPCs were then harvested after cell culture passage 5. They were subsequently transplanted within the sub-retinal space of 1 month old line-3 rats. Using the Zeiss laser microdissection system, the transplanted layer of RPCs were gathered. Total RNA was purified (RNeasy Micro Kit, Qiagen, Valencia, Ca) from RPCs prior to and after transplantation. Gene expression patterns were compared between pre- and post-transplant RPCs with a rat genome microarray (Rat Genome 230 2.0 Array, Affymetrix Inc., Santa Clara, Ca). Data analysis was performed using the S-score algorithm to determine statistical significance. The Gene Ontology Database was used to determine the biological significance of a selection of differentially expressed genes.
Setting a p-value < 10-2, genes were selected that were differentially expressed within pre and post-transplant RPCs. The following biological processes were enriched within the differentially expressed group of genes: (1) detection of light stimulus (9.23 fold); (2) regulation of exocytosis (7.48 fold); (3) gamma-aminobutyric acid signaling pathway (6.80 fold); (4) generation of a signal involved in cell-cell signaling (3.40 fold). As an example, genes involved in detection of light stimulus were found to be differentially expressed including: GNAT2, GRM6, SAG, RGR, GNGT2. Similarly, we discovered several genes that were differentially expressed within the processes mentioned above.
Differences in gene expression between retinal progenitor cells before and after transplantation reveal specific biological processes that differ once RPCs are transplanted within the subretinal space. These differences highlight the importance of the subretinal milieu in regulating gene expression within RPCs.
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