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(Investigative Ophthalmology and Visual Science. 2003;44:2211-2220.)
© 2003 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.02-0704
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Differential Temporal and Spatial Expression of Immediate Early Genes in Retinal Neurons after Ischemia–Reperfusion Injury

Nagahisa Yoshimura,1 Takanobu Kikuchi,2 Sachiko Kuroiwa,1 and Satoko Gaun1

1From the Department of Ophthalmology and the 2Research Center for Instrumental Analysis, Shinshu University School of Medicine, Matsumoto, Japan.

PURPOSE. To investigate genes that are up- and downregulated in rat retinal ischemia–reperfusion injury systematically by using an oligonucleotide microarray system and to determine temporal and spatial expression changes of some genes that showed upregulation in the analysis.

METHODS. Retinal ischemia was induced in rats by increasing intraocular pressure to 110 mm Hg for 1 hour. Gene expression at 12 hours after reperfusion was compared with that in the control retina by using oligonucleotide microarrays that display a total of 8800 genes and expressed sequence tags (ESTs). Temporal and spatial expression changes of immediate early genes and cell-cycle–related genes were studied by using real-time polymerase chain reaction (PCR) and immunohistochemical methods.

RESULTS. At 12 hours after reperfusion, 135 genes and ESTs were found to be up- or downregulated. The upregulated genes were classified into seven groups: (1) immediate early genes; (2) cell-cycle–related genes; (3) stress-responsive protein genes; (4) cell-signaling protein genes; (5) cell-adhesion and cell surface protein genes; (6) genes for translation and protein turnover; and (7) genes for metabolism. Real-time PCR analyses showed peaks of Fra-1 expression at 6 hours after reperfusion, whereas those for c-Jun, Jun B, and cyclin D1 were at 24 hours. Fra-1 and Jun B immunoreactivities were found in Müller cells, whereas c-Jun and cyclin D1 immunoreactivities were found in apoptotic retinal neurons.

CONCLUSIONS. Gene expression changes after a retinal ischemia–reperfusion injury were profiled by using an oligonucleotide microarray system. Seven groups of genes were found to be upregulated by the injury. Among the immediate early genes, Fra-1 and Jun B immunoreactivities were found in Müller cells whereas c-Jun and cyclin D1 immunoreactivities were found in apoptotic retinal neurons.




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