Retinal cell responses to elevated intraocular pressure

A gene array comparison between the whole retina and retinal ganglion cell layer

Ying Guo, William O. Cepurna, Jennifer A. Dyck, Tom A. Doser, Elaine Johnson, John Morrison

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

PURPOSE. To determine and compare gene expression patterns in the whole retina and retinal ganglion cell layer (RGCL) in a rodent glaucoma model. METHODS. IOP was unilaterally elevated in Brown Norway rats (N = 26) by injection of hypertonic saline and monitored for 5 weeks. A cDNA microarray was used on whole retinas from one group of eyes with extensive optic nerve injury and on RGCL isolated by laser capture microdissection (LCM) from another group with comparable injury, to determine the significantly up- or downregulated genes and gene categories in both groups. Expression changes of selected genes were examined by quantitative reverse transcription-PCR (qPCR) to verify microarray results. RESULTS. Microarray analysis of the whole retina identified 632 genes with significantly changed expression (335 up, 297 down), associated with 9 upregulated and 3 downregulated biological processes. In contrast, the RGCL microarray yielded 3726 genes with significantly changed expression (2003 up, 1723 down), including 60% of those found in whole retina. Thirteen distinct upregulated biological processes were identified in the RGCL, dominated by protein synthesis. Among 11 downregulated processes, axon extension and dendrite morphogenesis and generation of precursor metabolism and energy were uniquely identified in the RGCL. qPCR confirmed significant changes in 6 selected messages in whole retina and 11 in RGCL. Increased Atf3, the most upregulated gene in the RGCL, was confirmed by immunohistochemistry of RGCs. CONCLUSIONS. Isolation of RGCL by LCM allows a more refined detection of gene response to elevated pressure and improves the potential of determining cellular mechanisms in RGCs and their supporting cells that could be targets for enhancing RGC survival.

Original languageEnglish (US)
Pages (from-to)3003-3018
Number of pages16
JournalInvestigative Ophthalmology and Visual Science
Volume51
Issue number6
DOIs
StatePublished - Jun 2010

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Retinal Ganglion Cells
Intraocular Pressure
Retina
Genes
Laser Capture Microdissection
Biological Phenomena
Down-Regulation
Optic Nerve Injuries
Microarray Analysis
Dendrites
Oligonucleotide Array Sequence Analysis
Morphogenesis
Glaucoma
Energy Metabolism
Reverse Transcription
Axons
Rodentia
Immunohistochemistry
Gene Expression
Pressure

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience
  • Medicine(all)

Cite this

Retinal cell responses to elevated intraocular pressure : A gene array comparison between the whole retina and retinal ganglion cell layer. / Guo, Ying; Cepurna, William O.; Dyck, Jennifer A.; Doser, Tom A.; Johnson, Elaine; Morrison, John.

In: Investigative Ophthalmology and Visual Science, Vol. 51, No. 6, 06.2010, p. 3003-3018.

Research output: Contribution to journalArticle

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