PURPOSE: To test whether Müller glial cells sense, and respond to, mechanical tension in the retina. METHODS: A device was designed to stretch the retina at right angles to its surface, across retinal layers. Pieces of retina were mounted between two hollow tubes, and uniaxial force was applied to the tissue using a micrometer-stepping motor. Müller cells were selectively stained with the fluorescent, calcium-sensitive dye X-Rhod-1 and were monitored in real time during retinal stretch in vitro. Immunohistochemistry was used to study protein levels and activation of intracellular pathways in stretched retinas. RESULTS: Müller cells responded acutely with transient increases in fluorescence during stretch, indicative of increased intracellular calcium levels. All the Müller cells elongated uniformly, and there was no apparent difference between retinal layers in resistance against mechanical deformation. After stretch, Müller cells showed fast activation of extracellular signal-regulated kinase (after 15 minutes), upregulation of transcription factor c-Fos (after 1 hour), and basic fibroblast growth factor (after 3 hours). No changes in intermediate filament protein expression were observed in Müller cells up to 3 hours after stretch. CONCLUSIONS: A novel technique was developed for real-time monitoring of Müller cells during retinal stretch, which allowed the identification of Müller cells as a mechanoresponsive cell type. Mechanical stress triggers molecular responses in Müller cells that could prevent retinal damage.
PURPOSE: To test whether Müller glial cells sense, and respond to, mechanical tension in the retina. METHODS: A device was designed to stretch the retina at right angles to its surface, across retinal layers. Pieces of retina were mounted between two hollow tubes, and uniaxial force was applied to the tissue using a micrometer-stepping motor. Müller cells were selectively stained with the fluorescent, calcium-sensitive dye X-Rhod-1 and were monitored in real time during retinal stretch in vitro. Immunohistochemistry was used to study protein levels and activation of intracellular pathways in stretched retinas. RESULTS: Müller cells responded acutely with transient increases in fluorescence during stretch, indicative of increased intracellular calcium levels. All the Müller cells elongated uniformly, and there was no apparent difference between retinal layers in resistance against mechanical deformation. After stretch, Müller cells showed fast activation of extracellular signal-regulated kinase (after 15 minutes), upregulation of transcription factor c-Fos (after 1 hour), and basic fibroblast growth factor (after 3 hours). No changes in intermediate filament protein expression were observed in Müller cells up to 3 hours after stretch. CONCLUSIONS: A novel technique was developed for real-time monitoring of Müller cells during retinal stretch, which allowed the identification of Müller cells as a mechanoresponsive cell type. Mechanical stress triggers molecular responses in Müller cells that could prevent retinal damage.
Authors: David P Allison; Claretta J Sullivan; Ninell Pollas Mortensen; Scott T Retterer; Mitchel Doktycz Journal: J Vis Exp Date: 2011-08-10 Impact factor: 1.355
Authors: Daniel A Ryskamp; Andrew O Jo; Amber M Frye; Felix Vazquez-Chona; Nanna MacAulay; Wallace B Thoreson; David Križaj Journal: J Neurosci Date: 2014-11-19 Impact factor: 6.167
Authors: Dimitra Athanasiou; Monica Aguilà; Dalila Bevilacqua; Sergey S Novoselov; David A Parfitt; Michael E Cheetham Journal: FEBS Lett Date: 2013-05-15 Impact factor: 4.124