Abstract
Abstract: :
Purpose: Scleral fibroblasts are involved in scleral remodeling during axial elongation in myopia. Mechanical load is a potent stimulator of gene expression. This study seeks to identify changes in gene expression of scleral fibroblasts in response to mechanical load and possible mechanisms of scleral remodeling in the development of myopia. Methods: Human fibroblasts were mechanically stretched for 30 minutes and 24 hours. A gene expression microarray containing probes for a total of 12,000 genes was used to measure changes in gene expression in response to the induced strain. Results: 237 genes revealed differential and significant changes in expression (P<0.01) after 30 minutes of stretching, 28 unexpressed genes began to be expressed (turn on) in response to strain,while 31 expressed genes were no longer expressed (turn off). After24 hours of stretching, 308 genes showed reproducible changes in expression (P<0.01), 29 gene genes were turned on and 17 genes were turned off. After 30 minutes, 25 genes showed at least a threefold changes in expression. These included cell receptors (5 genes),protein kinases (5 genes),cell growth/differentiation factors (3 genes),Extracellular matrix (ECM) (3 genes), lipid metabolism (3 genes),protein metabolism (2 genes), transcription factors (2 genes), binding proteins (1 gene), and water channel (1 gene). After 24 hours,21 gene showed at least a threefold change in expression. These included cell receptors (6 genes), protein kinases (5 genes), cell growth/differentiation factors (4 genes),lipid metabolism (3 genes), ECM (1 genes), transcription factor (1 genes) and carbohydrate metabolism (1 gene). Two techniques (RT-PCR and Southern blotting) confirmed the changes in expression of scleral genes. Conclusions: We identified a large number of early and late mechanical response genes in human scleral fibroblasts.These changes in gene expression will provide potential candidate genes that may be involved in scleral remodeling during axial elongation in myopia. CR:None Support:NIH grants EY03040 and an unrestricted grant from Research to Prevent Blindness, New York,NY
Keywords: gene microarray • molecular biology