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Exploring phospho‐signaling pathways in patients with agerelated macular degeneration (AMD), human pigment epithelial cell line ARPE‐19 and porcine ocular tissue
Author(s) -
FaslerKan Elizaveta,
Barteneva Natasha,
Meyer Peter
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.645.11
Subject(s) - flow cytometry , retina , microbiology and biotechnology , retinal pigment epithelium , retinal , macular degeneration , signal transduction , cell culture , confocal microscopy , downregulation and upregulation , biology , pathology , chemistry , cancer research , medicine , ophthalmology , biochemistry , gene , genetics , neuroscience
The pig has been used extensively in experimental studies of human eye diseases. Pig retina shares many morphological features with the human eye. Retinal pigment epithelial cells constitute an important component of the blood‐retinal barrier and play a pivotal role in the development of AMD. Understanding the underlying molecular mechanisms is a prerequisite for developing therapeutic strategies. We investigated cytokine‐induced changes of the JAK/STAT and NF‐kappa B signaling pathways. Methods Porcine ocular cells, human ARPE‐19 cell line, eyes with AMD were used to study the nuclear translocation of STAT and NF‐kB proteins by gel‐shift assay, immunohistochemistry, confocal laser microscopy and flow cytometry. Results Histological examination of AMD patients revealed a strong formation of choroidal neovascular membranes in the macular region. Strong activation of STAT3 was observed in these membranes. Strong STAT1 activation was demonstrated upon exposure to interferon‐γ in ARPE‐19 and pig ocular cells in vitro . Human recombinant interferon‐α upregulated STAT1, STAT2 and STAT3, while IL‐6 and IL‐4 activated STAT3 and STAT6, respectively. Confocal microscopy identified the nuclear translocation of the STAT proteins. Conclusion Our data demonstrate the upregulation of the members of the JAK‐STAT signaling pathway in AMD patients, porcine ocular cells and ARPE‐19 cell line.