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LK Geiger, JR Mertz; Differentially Expressed Chick Retinal and Choroidal Genes During Form Deprivation Myopia And Recovery From Myopia . Invest. Ophthalmol. Vis. Sci. 2002;43(13):204.
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Purpose: Form deprivation (FD) causes increases in eye growth rate in chick eyes. Conversely, recovery from FD attenuates the rate of eye growth. We employed a PCR-based subtractive hybridization technique to isolate clones of differentially expressed genes that may induce the changes observed in FD and recovery from FD. Methods: Eleven 3-day-old chicks were deprived of form vision with translucent occluders for 13 days; five chicks recovered for 3 days without occluders, and the other six chicks remained occluded for 3 days. Changes in eye growth and choroidal thickness were determined using high-frequency A scan ultrasonography. Total RNA was isolated from retinal and choroidal tissue. The RNA was reverse-transcribed and used as templates for subtractive hybridization using a variation of the Clontech PCR-Select Differential Screening Kit, in which biotin was incorporated in the subtracter. After subtractive hybridization, gel products were purified and cloned. Results: We observed upregulation of approximately 5 clones in retinas from FD eyes compared to eyes that recovered; in addition, we found 6 upregulated clones in retinas from eyes that recovered from FD compared to the FD retinas. For choroids, only the eyes that were FD demonstrated upregulation of approximately 4 clones. Conclusion: To our knowledge, this is the first time PCR-based subtractive hybridization has been used to examine gene expression in chick eyes with experimentally-induced changes in eye growth. This technique isolated potential gene clones that may be modulated by visual experience. We are in the process of sequencing and identifying these clones.
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