Differential temporal and spatial expression of immediate early genes in retinal neurons after ischemia-reperfusion injury.
OBJECTIVE
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.
Yoshimura
Nagahisa
N
Department of Ophthalmology, Shinshu University School of Medicine, Matsumoto, Japan. nagaeye@hsp.md.shinshu-u.ac.jp
Kikuchi
Takanobu
T
Kuroiwa
Sachiko
S
Gaun
Satoko
S
D000818:Animals; D018797:Cell Cycle Proteins / Q000235:genetics; D005136:Eye Proteins / Q000235:genetics; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D020869:Gene Expression Profiling; D017781:Genes, Immediate-Early / Q000235:genetics*; D017874:Immediate-Early Proteins / Q000235:genetics; D008297:Male; D009474:Neurons / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000378:metabolism*; D012160:Retina / Q000378:metabolism*; D012164:Retinal Diseases / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation
D018797:Cell Cycle Proteins; D005136:Eye Proteins; D017874:Immediate-Early Proteins; D012333:RNA, Messenger