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Elie Beit-Yannai, Natali Lerner; Gamma-catenin is presented in non-pigmented ciliary epithelium derived exosomes, suggest downstream roll in the drainage system homeostasis. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):3262.
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Exosomes within the aqueous humor are part of the drainage system homeostasis. Non-pigmented ciliary epithelium (NPCE) derived exosome can contribute to this homeostasis. In this study, we characterized NPCE derived exosomes and identified major protein content.
NPCE cell line (ODM-2) in monolayer, 90% confluent, was grown for 48h in exosome depleted media. The condition media exosomes were extracted by ultracentrifugation and the exosomes were characterized by: dynamic light scattering, zeta potential, sucrose gradient, cryo-electron microscopy, western blot, 2Dgell electrophoresis and MS.
Results show that NPCE derived exosomes size are 101±13.2nm, has a zeta potential of -20.9±6mV according to DLS analysis. Cryo-TEM analysis demonstrated cell size variations 40-130nm. By sucrose gradient exosomes were found to have density of 1.13-1.19 g/ml with high expression of the marker proteins Alix and Tsg 101. 2D gel electrophoresis demonstrated a high concentration of gamma-catenine.
NPCE cells derived exosomes suggest a source for drainage system regulating proteins such as gamma-catenine who mediate mediates the cross-talk between cadherin-based adhesion, catenin-dependent transcription and Wnt signaling.
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