PMID: 11923227Mar 30, 2002Paper

Extracellular signal-regulated kinase activation predominantly in Müller cells of retina with endotoxin-induced uveitis

Investigative Ophthalmology & Visual Science
Masumi TakedaH Kiyama

Abstract

To evaluate the consequences of mitogen-activated protein kinase (MAPK)-mediated signaling in retinas with endotoxin-induced uveitis (EIU). EIU was induced with footpad inoculation of lipopolysaccharide (LPS). To identify the expression and activity of extracellular signal-regulated kinase (ERK), c-Jun NH2-terminal kinase (JNK), and p38, Western blot analysis and immunohistochemistry (IHC) were performed using antibodies against these kinases and phosphorylated forms. To evaluate the ERK mRNA expression level, semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was performed. To identify cell species that express phosphorylated (p)-ERK, simultaneous demonstration of p-ERK and glial fibrillary acidic protein (GFAP) was performed with combined IHC and in situ hybridization. Dexamethasone (Dex) was used to reduce the LPS-induced inflammatory stimulus, and changes in p-ERK expression were evaluated by Western blot analysis after treatment. Only p-ERK among the phosphorylated MAPKs increased after LPS stimulation, according to Western blot analysis. p-ERK increased after LPS injection, whereas both the Western blot and RT-PCR studies showed no apparent changes in ERK-1 and -2 expression. IHC revealed that stron...Continue Reading

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