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J. Zhou, E. Rappaport, T.L. Young; Microarray Analysis of Gene Expression in Mouse Sclera . Invest. Ophthalmol. Vis. Sci. 2005;46(13):3332.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To develop gene expression profiles in mouse sclera to allow for the identification of novel, uncharacterized genes in this tissue–type, and to identify candidate genes for scleral disorders. Methods: Eighteen adult C57BL/6J mice were sacrificed at 8 weeks of age. Mouse sclera total RNA was isolated from 3 groups of mice (6 per group) using a standard TriZol reagent protocol, and reverse transcribed into cDNA using the Invitrogen Superscript cDNA synthesis kit. The resulting cDNA was in vitro transcribed to produce biotin–labeled cRNA, fragmented into 50 – 200 base pair products, mixed with hybridization controls before a 16 hour hybridization step with oligonucleotide probes, and hybridized to 3 Affymetrix GeneChip® Mouse Genome 430 2.0 Array chips. Data collection and array analyses were carried out with GeneChip Operating Software version 1.2 (Affymetrix), using the expression analysis algorithm to run an absolute analysis after cell intensities were computed. All arrays were scaled to the same target intensity using all probe sets. Results: There were 10,466 genes from over 39,000 gene chip transcripts with "present" calls assigned independently to each of the 3 mouse sclera groups. These genes could be clustered into 6 major categories: physiological process (32.9%); cellular process (20.1%); extracellular components (9.3%); signal transducer activity (5.3%); development (4.8%); and transcription regulator activity (3.4%). Conclusions: A comprehensive gene expression profile of normal mouse sclera has been determined. This work may aid in animal–model studies of eye diseases involving changes in scleral physiology.
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