Abstract
Purpose:
To determine gene expression in the corneal endothelium of healthy children and adults to provide insight into the normal age-associated changes in corneal endothelial cell gene expression.
Methods:
Total RNA was isolated from corneal endothelium obtained from four pediatric (≤11 years old) and four adult (≥53 years old) donor eye bank corneas. The RNA samples were processed and hybridized to the Affymetrix GeneChip 1.1ST array. Analysis of the raw intensity values contained within CEL files was performed using the gene expression module within the Partek Genomics Suite software. A list of the most significant differentially expressed genes between pediatric and adult corneal endothelium was obtained using a p-value and fold-change cutoff of 0.01 and 2, respectively.
Results:
Principal component analysis (PCA) of our expression dataset demonstrated two distinct populations of samples in PCA1 with each being comprised of either the pediatric or adult samples, while hierarchical clustering showed a strong relationship within samples originating from the same age group. Differential gene expression analysis identified ten significantly regulated genes, of which seven encode for protein: ITGBL1, PREX2, DIO2, DLL4, C9orf131, HIST1H3A, and TNFAIP3.
Conclusions:
The identification of several differentially expressed genes in pediatric and adult corneal endothelial cells suggests that changes in gene transcription continue well after birth. After validating the results of this study, we plan to investigate the role of each significantly differentially expressed gene in normal and abnormal corneal endothelial cell function.
Keywords: 481 cornea: endothelium •
533 gene/expression •
535 gene microarray