May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Regulation of Extracellular Myocilin Associated Exosomes from Trabecular Meshwork Cells
Author Affiliations & Notes
  • E. A. Hoffman
    University of Arizona, Tucson, Arizona
    Ophthalmology,
  • K. M. Perkumas
    University of Arizona, Tucson, Arizona
    Ophthalmology,
  • K. M. Hardy
    University of Arizona, Tucson, Arizona
    Physiology,
  • W. D. Stamer
    University of Arizona, Tucson, Arizona
    Ophthalmology,
  • Footnotes
    Commercial Relationships E.A. Hoffman, None; K.M. Perkumas, None; K.M. Hardy, None; W.D. Stamer, None.
  • Footnotes
    Support EY12797 (WDS), RPB (WDS)
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 2063. doi:https://doi.org/
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    • Get Citation

      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)

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Abstract

Purpose:: 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.

Methods:: 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.

Results:: 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.

Conclusions:: Both dexamethasone and γ-interferon significantly upregulate the expression of MYOC and its association with exosomes in conditioned media from TM cells.

Keywords: trabecular meshwork • proteins encoded by disease genes • protein purification and characterization 
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