Retinal gene expression after central retinal artery ligation: effects of ischemia and reperfusion.
OBJECTIVE
To investigate the morphologic and molecular consequences of 30- and 90-minute central retinal artery ligation (CRAL)-induced retinal ischemia, followed by 3 and 12 hours of reperfusion, and to identify potential targets for therapy.


METHODS
Retinal ischemia was induced for 30 and 90 minutes by ligating the rat central retinal artery, and corresponding effects were examined histologically, immunocytochemically, and molecularly at 3 hours and 12 hours of reperfusion. Patterns of gene expression revealed significantly upregulated and downregulated genes by gene array analyses and were verified by quantitative RT-PCR. Functional pathways were correlated using gene set enrichment analysis.


RESULTS
Substantial morphologic changes occurred from 3 hours to 7 days after CRAL in rats, resulting in a cellular loss in most retinal layers, particularly in inner nuclear and ganglion cell layers. After 30 minutes of CRAL and 3 hours of reperfusion, transcription-related genes such as ATF3, ID2, Klf4, BTG2, c-Fos, and c-Jun were activated. After 12 hours of reperfusion, the genes associated with kinase and caspase molecular pathways-including MAP kinases, Casp3 and Casp9-were upregulated. CRAL of 90 minutes and 3 hours of reperfusion induced glycolysis and gluconeogenesis-related genes such as G6PC. However, prolonged reperfusion of 12 hours was characterized by prominent activation of apoptosis-related genes, including Tp53, Akt1, Akt3, Pik3R1, Prkcb1, caspases (Casp3, Casp7, Casp9), and TNF.


CONCLUSIONS
CRAL is a clinically relevant retinal ischemia model, and gene expression analysis can provide information regarding the molecular mechanisms underlying the pathophysiological processes during retinal ischemia. In addition, CRAL represents an effective experimental model with which to study retinal inflammation, development, aging, and, neurodegeneration.
Prasad
Shiv S
SS
Genomics Division, Biologics and Genetic Therapies Directorate, Health Canada, Ottawa, Ontario, Canada. shiv_prasad@hc-sc.gc.ca
Kojic
Ljubomir
L
Wen
Yan H
YH
Chen
Zheng
Z
Xiong
Wei
W
Jia
William
W
Cynader
Max S
MS
D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000235:genetics; D005136:Eye Proteins / Q000235:genetics*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology*; D020287:In Situ Nick-End Labeling; D007511:Ischemia / Q000235:genetics* / Q000378:metabolism / Q000503:physiopathology; D000090062:Kruppel-Like Factor 4; D008026:Ligation; D008297:Male; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D051381:Rats; D011917:Rats, Inbred Lew; D015427:Reperfusion Injury / Q000235:genetics* / Q000378:metabolism / Q000503:physiopathology; D012160:Retina / Q000378:metabolism*; D012161:Retinal Artery / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics
D051017:Apoptosis Regulatory Proteins; D005136:Eye Proteins; C000715469:Klf4 protein, rat; D000090062:Kruppel-Like Factor 4; D014157:Transcription Factors