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The Role of Complement and the Extracellular Matrix in Early Stage Macular Degeneration
Author(s) -
FernandezGodino Rosario,
Pierce Eric,
Garland Donita
Publication year - 2015
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.29.1_supplement.719.21
Subject(s) - microbiology and biotechnology , extracellular matrix , extracellular , complement system , biology , phenotype , mutant , immunology , antibody , gene , genetics
Gene targeted Efemp1 R345W/R345W knockin mice are a documented model of the Efemp1 ‐associated macular degeneration. These mice develop features of macular degenerations including extensive basal deposits when the complement system is active. The aim of this project was to use cultured primary RPE cells to study the mechanisms involved in the pathogenesis of basal deposit formation and the role of the complement system. Mice were used in conformance with the FASEB Statement of Principles for the use of animals in research and education. Primary RPE cells from wild type, Efemp1 R345W/R345W , and Efemp1 R345W/R345W : C3 ‐/‐ mice were grown on transwells. Cell phenotypes were characterized by electron microscopy. The presence of complement components and factors, extracellular matrix proteins and pro‐inflammatory molecules were quantified by qRT‐PCR, immunofluorescence and ELISA. The activities of specific proteins were modulated with antibodies and recombinant proteins. We conclude that cultured primary RPE cells from Efemp1 R345W/R345W knockin but not from wild type or double mutant Efemp1 R345W/R345W : C3 ‐/‐ mice form extracellular deposits. The deposits contain extracellular matrix and complement proteins. The formation of deposits can be inhibited by neutralizing C3a, IL‐1B, IL‐6 or EFEMP1 in Efemp1 R345W/R345W cultures. Wild type cells make deposits in the presence of C3a or recombinant mutant EFEMP1. We conclude that RPE cells from Efemp1 R345W/R345W knockin mice generate the extracellular deposits through a C3a dependent mechanism that is activated by EFEMP1 R345W . These cells thus provide a model system to investigate the early mechanisms responsible for RPE dysfunction in macular degeneration.