DNA microarray analysis of gene expression in iris and ciliary body of rat eyes with endotoxin-induced uveitis.
The purposes of this study are to determine the genes that are up- or down-regulated in eyes with endotoxin-induced uveitis (EIU) by an oligonucleotide microarray system, and to determine the temporal and spatial changes in expression of selected genes that show strong up-regulation. EIU was induced by a footpad injection of lipopolysaccharide (LPS) in male Lewis rats. The expression of genes in the iris-ciliary body (ICB) at 2, 6, 12, and 24 hr after LPS injection was determined by oligonucleotide microarray analyses and compared to that in control rats. The microarray displayed 9911 genes and expressed sequence tags (ESTs). Cluster analysis was performed for highly up-regulated genes. Selected genes for cytokines (interleukin (IL)-1 beta and IL-6), chemokines (RANTES), and immediate early genes (Jun B, c-Fos, and c-Jun) were also studied by real-time polymerase chain reaction (PCR). Immunohistochemical studies were performed to localize the protein expression of some immediate early gene products. After LPS injection, the expression of 1930 genes were increased or decreased over 2-folds compared with normal controls by 24 hr. One hundred and seventeen genes were up-regulated over 10-fold, and these were classified into five clusters with similar expression pattern. The immediate early genes and transcription factors genes were included in one cluster of up-regulated genes peaking at 2 hr after the LPS injection. The expressions of cytokines, chemokines, and adhesion molecules were highly up-regulated. Real-time PCR analyses for selected genes showed similar expression changes as detected by the microarray analyses. Jun B immunoreactivity was found in the ICB cells at 3 and 6 hr after LPS injection. Gene expression changes after LPS injection were profiled by using an oligonucleotide microarray system. Our data suggest that the immediate early genes, such as Jun B, play an important role in inducing the inflammatory-related genes in the ICB.
Ohta
Kouichi
K
Department of Ophthalmology, School of Medicine, Shinshu University, 3-1-1 Asahi, Matsumoto 390-8621, Japan. kohta@hsp.md.shinshu-u.ac.jp
Kikuchi
Takanobu
T
Miyahara
Teruyoshi
T
Yoshimura
Nagahisa
N
D000818:Animals; D001082:Aqueous Humor / Q000737:chemistry / Q000166:cytology; D002924:Ciliary Body / Q000502:physiology*; D016000:Cluster Analysis; D015536:Down-Regulation / Q000235:genetics; D005136:Eye Proteins / Q000032:analysis; D005786:Gene Expression Regulation / Q000235:genetics*; D007150:Immunohistochemistry / Q000379:methods; D007498:Iris / Q000502:physiology*; D008070:Lipopolysaccharides; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011917:Rats, Inbred Lew; D013997:Time Factors; D015854:Up-Regulation / Q000235:genetics; D014605:Uveitis / Q000235:genetics*
D005136:Eye Proteins; D008070:Lipopolysaccharides; D012333:RNA, Messenger