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E. A. Hoffman, K. M. Perkumas, K. M. Hardy, W. D. Stamer; Regulation of Extracellular Myocilin Associated Exosomes from Trabecular Meshwork Cells. Invest. Ophthalmol. Vis. Sci. 2007;48(13):2063. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Myocilin (MYOC) is the protein product of a glaucoma-associated gene. MYOC appears extracellularly in vitro and in vivo in association with extracellular vesicles called exosomes, which function in the transport of lipid-associated ligands between cell neighbors. The goal of the present study was to determine ways to regulate the release of MYOC associated exosomes from trabecular meshwork (TM) cells.
Conditioned media from TM cells were analyzed for MYOC associated exosomes after treatment with dexamethasone (1µM) or γ-interferon (500 units/mL) for 5 days. Exosomes from conditioned medium were purified by differential centrifugation as described previously. Proteins in exosome and soluble fractions were separated by SDS-PAGE and analyzed for MYOC content by western blotting.
The appearance of extracellular MYOC in association with exosomes isolated from dexamethasone treated TM cell conditioned media increased by 261% and the total extracellular fraction increased 37.9%. Treatment with γ- interferon resulted in the appearance of the exosome marker HLA-DR and increased MYOC association with exosomes by 40.5% over untreated cells in culture. By comparison, total extracellular MYOC decreased by 29.5% upon γ- interferon treatment.
Both dexamethasone and γ-interferon significantly upregulate the expression of MYOC and its association with exosomes in conditioned media from TM cells.
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