Microarray analysis of retinal gene expression in the DBA/2J model of glaucoma

Michael R. Steele, Denise M. Inman, David J. Calkins, Philip J. Horner, Monica L. Vetter

Research output: Contribution to journalArticle

158 Citations (Scopus)

Abstract

PURPOSE. The DBA/2J mouse is a model for secondary angle-closure glaucoma, due to iris atrophy and pigment dispersion, which ultimately lead to increased intraocular pressure (IOP). The study was undertaken to correlate changes in retinal gene expression with IOP elevation by performing microarray analysis of retinal RNA from DBA/2J mice at 3 months before disease onset and at 8 months after IOP elevation. METHODS. IOP was monitored monthly in DBA/2J animals, and animals with normal (3 months) or elevated IOP (8 months) were identified. RNA was prepared from three individual retinas at each age, and the RNA was amplified and used to generate biotin-labeled probe for high-density mouse gene microarrays (U430.2; Affymetrix, Santa Clara, CA). A subset of genes was selected for confirmation by quantitative RT-PCR, by using independent retina samples from DBA/2J animals at 3, 5, and 8 months of age and compared to retinas from C57BL/6J control animals at 3 and 8 months. RESULTS. There were changes in expression of 68 genes, with 32 genes increasing and 36 genes decreasing at 8 months versus 3 months. Upregulated genes were associated with immune response, glial activation, signaling, and gene expression, whereas downregulated genes included multiple crystallin genes. Significant changes in nine upregulated genes and two downregulated genes were confirmed by quantitative RTPCR, with some showing changes in expression by 5 months. CONCLUSIONS. DBA/2J retina shows evidence of glial activation and an immune-related response after IOP elevation, similar to what has been reported after acute elevation of IOP in other models.

Original languageEnglish
Pages (from-to)977-985
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume47
Issue number3
DOIs
StatePublished - 1 Mar 2006

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Microarray Analysis
Glaucoma
Intraocular Pressure
Gene Expression
Genes
Retina
Inbred DBA Mouse
RNA
Neuroglia
Down-Regulation
Angle Closure Glaucoma
Crystallins
Iris
Biotin
Atrophy
Polymerase Chain Reaction

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Steele, Michael R. ; Inman, Denise M. ; Calkins, David J. ; Horner, Philip J. ; Vetter, Monica L. / Microarray analysis of retinal gene expression in the DBA/2J model of glaucoma. In: Investigative Ophthalmology and Visual Science. 2006 ; Vol. 47, No. 3. pp. 977-985.
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Microarray analysis of retinal gene expression in the DBA/2J model of glaucoma. / Steele, Michael R.; Inman, Denise M.; Calkins, David J.; Horner, Philip J.; Vetter, Monica L.

In: Investigative Ophthalmology and Visual Science, Vol. 47, No. 3, 01.03.2006, p. 977-985.

Research output: Contribution to journalArticle

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T1 - Microarray analysis of retinal gene expression in the DBA/2J model of glaucoma

AU - Steele, Michael R.

AU - Inman, Denise M.

AU - Calkins, David J.

AU - Horner, Philip J.

AU - Vetter, Monica L.

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N2 - PURPOSE. The DBA/2J mouse is a model for secondary angle-closure glaucoma, due to iris atrophy and pigment dispersion, which ultimately lead to increased intraocular pressure (IOP). The study was undertaken to correlate changes in retinal gene expression with IOP elevation by performing microarray analysis of retinal RNA from DBA/2J mice at 3 months before disease onset and at 8 months after IOP elevation. METHODS. IOP was monitored monthly in DBA/2J animals, and animals with normal (3 months) or elevated IOP (8 months) were identified. RNA was prepared from three individual retinas at each age, and the RNA was amplified and used to generate biotin-labeled probe for high-density mouse gene microarrays (U430.2; Affymetrix, Santa Clara, CA). A subset of genes was selected for confirmation by quantitative RT-PCR, by using independent retina samples from DBA/2J animals at 3, 5, and 8 months of age and compared to retinas from C57BL/6J control animals at 3 and 8 months. RESULTS. There were changes in expression of 68 genes, with 32 genes increasing and 36 genes decreasing at 8 months versus 3 months. Upregulated genes were associated with immune response, glial activation, signaling, and gene expression, whereas downregulated genes included multiple crystallin genes. Significant changes in nine upregulated genes and two downregulated genes were confirmed by quantitative RTPCR, with some showing changes in expression by 5 months. CONCLUSIONS. DBA/2J retina shows evidence of glial activation and an immune-related response after IOP elevation, similar to what has been reported after acute elevation of IOP in other models.

AB - PURPOSE. The DBA/2J mouse is a model for secondary angle-closure glaucoma, due to iris atrophy and pigment dispersion, which ultimately lead to increased intraocular pressure (IOP). The study was undertaken to correlate changes in retinal gene expression with IOP elevation by performing microarray analysis of retinal RNA from DBA/2J mice at 3 months before disease onset and at 8 months after IOP elevation. METHODS. IOP was monitored monthly in DBA/2J animals, and animals with normal (3 months) or elevated IOP (8 months) were identified. RNA was prepared from three individual retinas at each age, and the RNA was amplified and used to generate biotin-labeled probe for high-density mouse gene microarrays (U430.2; Affymetrix, Santa Clara, CA). A subset of genes was selected for confirmation by quantitative RT-PCR, by using independent retina samples from DBA/2J animals at 3, 5, and 8 months of age and compared to retinas from C57BL/6J control animals at 3 and 8 months. RESULTS. There were changes in expression of 68 genes, with 32 genes increasing and 36 genes decreasing at 8 months versus 3 months. Upregulated genes were associated with immune response, glial activation, signaling, and gene expression, whereas downregulated genes included multiple crystallin genes. Significant changes in nine upregulated genes and two downregulated genes were confirmed by quantitative RTPCR, with some showing changes in expression by 5 months. CONCLUSIONS. DBA/2J retina shows evidence of glial activation and an immune-related response after IOP elevation, similar to what has been reported after acute elevation of IOP in other models.

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