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M Tsujikawa, K Tsujikawa, S Matsuda, F Gomi, Y Tano; Comprehensive Gene Expression Analysis of Human Neurosensory Retina . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3656.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To investigate the gene expression profile of the human neurosensory retina. Methods: We performed serial analysis of gene expression (SAGE), which allows quantitative and comprehensive analysis of gene expression. A SAGE library was constructed from 40 ug of human retina total RNA and sequenced. The sequence data were analyzed by SAGE software and matched to GenBank and UniGene public databases. The expression patterns was compared with that of human brain, and according to the some tags highly expressed only in retina, the genes corresponding to the tags and which expression patterns were confirmed by 3*fRACE and RT-PCR, respectively. Results: To date, we have analyzed ∼25000 tags were, and in which ∼11000 of which were unique. The tags that were highly expressed at levels greater than 0.1% of transcript accounted for ∼30% of the transcript mass, however, this accounted for ∼2.0% of the total number of unique genes. The expression of only five retina-specific genes (rod outer segment membrane protein 1 [ROM1], retinal photoreceptor synaptic protein [HRG4], cGMP phosphodiesterase gamma-subunit [PDEG], guanylate kinase [GUK1], and arrestin) constituted ∼3.0% of the total transcript mass. The expression level of glutathione peroxidase 3 was 0.67% of the transcript mass, and 100-fold greater than its expression level in the brain. The expression levels of the these genes were confirmed by RT-PCR. Conclusion: The advantage of SAGE over other methods of gene expression analysis is the high throughput that can be achieved. The expression profile of the normal retina presented here could be used as baseline data for future studies of differentially expressed genes in the developmental stage, genotype, the disease state, and drug and growth factor treatment.
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