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Kenneth Olsen, John Flanagan; The Biomechanical Response of Optic Nerve Head Astrocytes to Compressive Insult. Invest. Ophthalmol. Vis. Sci. 2013;54(15):72.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the biomechanical response of human optic nerve head astrocytes to compressive insult.
Primary human optic nerve head astrocytes were grown to confluence on pre-stretched, sylastic membranes (Flexcell® Bioflex® plates), using a custom built device. Media was changed twice weekly with DMEM/F12 media containing 10% FBS and 1% pen/strep. Upon reaching confluence the cells were serum deprived for 24 hours and subjected to 12% cyclic compression at 1 Hz using the FlexerCell® Strain System FX-4000 for 2 hours (n=3). Control cells were also serum deprived for 24 hours followed by another 2 hours (n=3). Cell lysates were collected and proteomic analysis was carried out to determine protein regulation using the LTQ Orbitrap Velos®. Proteins of interest were selected based on statistical significance (p<0.05) and fold change (>±1.5) and analyzed using the Ingenuity Systems' IPA® biomolecular analysis software.
Of the 875 unique proteins discovered 60 were determined to play an important role in the response of astrocytes to compressive strain. Several networks were identified to be involved in the response including cell morphology, cellular assembly and organization, cellular function and maintenance, cell cycle, and cell-to-cell signaling and interaction. Proteins of particular interest for further mechanistic investigation included FLNB (+4.3 fold), TGFBI (+4.1 fold), and RHOA (+1.6). Proteins previously identified to be regulated by biomechanical stretch were also seen here including ANXA4 (-2.1 fold), S100 (A16, -10.0 fold), and ROA (+1.7 fold).
We have identified several proteins and protein networks of interest following the response of human ONH astrocytes to compressive insult. In particular several proteins involved in pathways known to be regulated in activated astrocytes were identified including TGFBI which is associated with extra cellular matrix remodelling, and RHOA and FLNB which are involved with reorganization of the cytoskeleton and cell morphology.
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