PURPOSE: To investigate changes in retinal ganglion cell (RGC) survival and morphology, retinal glial cell effects on RGC survival, and changes in mRNA expression during centrifugal force loading using a newly developed device. METHODS: Changes in RGC survival and morphology were examined when isolated RGCs from 2-day-old rats were loaded with centrifugal force equivalent to 16, 28, or 33 mm Hg. The effects of cocultured retinal glial cells on RGC survival were studied in the presence of centrifugal force equivalent to 16 and 28 mm Hg for 48 hours. The microarray method and real-time polymerase chain reaction confirmed changes in mRNA expression when RGCs and retinal glial cells were loaded with centrifugal force equivalent to 28 mm Hg for 24 hours. RESULTS: The survival of isolated RGCs and the number of neurites were significantly decreased by centrifugal force loading. Conversely, there was no significant change in the survival of isolated retinal glial cells. The survival of cocultured RGCs was significantly better than that of isolated RGCs. In contrast to the numerous changes in the mRNA expression of retinal glial cells subjected to centrifugal force loading, there was no significant change in the mRNA expression of RGCs. CONCLUSIONS: The developed device may have potential for use as an in vitro model of RGC damage. The response to centrifugal force loading varies according to cell type, and the marked changes in the mRNA expression of retinal glial cells may be involved in the improvement of RGC survival.
PURPOSE: To investigate changes in retinal ganglion cell (RGC) survival and morphology, retinal glial cell effects on RGC survival, and changes in mRNA expression during centrifugal force loading using a newly developed device. METHODS: Changes in RGC survival and morphology were examined when isolated RGCs from 2-day-old rats were loaded with centrifugal force equivalent to 16, 28, or 33 mm Hg. The effects of cocultured retinal glial cells on RGC survival were studied in the presence of centrifugal force equivalent to 16 and 28 mm Hg for 48 hours. The microarray method and real-time polymerase chain reaction confirmed changes in mRNA expression when RGCs and retinal glial cells were loaded with centrifugal force equivalent to 28 mm Hg for 24 hours. RESULTS: The survival of isolated RGCs and the number of neurites were significantly decreased by centrifugal force loading. Conversely, there was no significant change in the survival of isolated retinal glial cells. The survival of cocultured RGCs was significantly better than that of isolated RGCs. In contrast to the numerous changes in the mRNA expression of retinal glial cells subjected to centrifugal force loading, there was no significant change in the mRNA expression of RGCs. CONCLUSIONS: The developed device may have potential for use as an in vitro model of RGC damage. The response to centrifugal force loading varies according to cell type, and the marked changes in the mRNA expression of retinal glial cells may be involved in the improvement of RGC survival.
Authors: Mabel Enriquez-Algeciras; Di Ding; Fabrizio G Mastronardi; Robert E Marc; Vittorio Porciatti; Sanjoy K Bhattacharya Journal: J Clin Invest Date: 2013-01-02 Impact factor: 14.808