PURPOSE: To investigate whether cell cycle-related genes play a role in neuronal cell death in retinal ischemia-reperfusion injury. METHODS: Retinal ischemia-reperfusion injury was induced in rats by a ligation method and also by increasing the intraocular pressure. After 1 hour-of ischemia, cell death in the retina was studied using the TdT-dUTP terminal nick-end labeling (TUNEL) method, propidium iodide (PI) staining, DNA ladder formation, and ultrastructural studies. Immunohistochemical studies using antibodies against cell cycle-related genes were conducted. Changes in expression of cyclin D1 mRNA were quantitated using competitive quantitative polymerase chain reaction. RESULTS: At 3 hours after reperfusion, cells in the ganglion cell layer were the first to die, followed by those in the inner nuclear layer (at 6 hours) and outer nuclear layer (at 9 hours). Ultrastructural studies revealed condensed nuclei and relatively preserved mitochondria; DNA ladder formation was also detected. Immunostaining was positive for the cell cycle-related gene products c-Jun, cyclin B1, and cyclin D1. The time course of TUNEL-positive cells and that of cells positive for c-Jun or cyclin D1 in the inner nuclear layer was similar. A double-labeling study, using PI or TUNEL, and immunohistochemical analysis revealed that dying cells expressed c-Jun and cyclin D1, whereas cyclin B1 expression was observed in Müller cells. Quantitation of cyclin D1 mRNA revealed an approximate 4-fold increase at 24 hours after reperfusion. CONCLUSIONS: Aberrant expression of cell cycle-related genes may play an important role in the cell death that accompanies retinal ischemia-reperfusion injury.
PURPOSE: To investigate whether cell cycle-related genes play a role in neuronal cell death in retinal ischemia-reperfusion injury. METHODS:Retinal ischemia-reperfusion injury was induced in rats by a ligation method and also by increasing the intraocular pressure. After 1 hour-of ischemia, cell death in the retina was studied using the TdT-dUTP terminal nick-end labeling (TUNEL) method, propidium iodide (PI) staining, DNA ladder formation, and ultrastructural studies. Immunohistochemical studies using antibodies against cell cycle-related genes were conducted. Changes in expression of cyclin D1 mRNA were quantitated using competitive quantitative polymerase chain reaction. RESULTS: At 3 hours after reperfusion, cells in the ganglion cell layer were the first to die, followed by those in the inner nuclear layer (at 6 hours) and outer nuclear layer (at 9 hours). Ultrastructural studies revealed condensed nuclei and relatively preserved mitochondria; DNA ladder formation was also detected. Immunostaining was positive for the cell cycle-related gene products c-Jun, cyclin B1, and cyclin D1. The time course of TUNEL-positive cells and that of cells positive for c-Jun or cyclin D1 in the inner nuclear layer was similar. A double-labeling study, using PI or TUNEL, and immunohistochemical analysis revealed that dying cells expressed c-Jun and cyclin D1, whereas cyclin B1 expression was observed in Müller cells. Quantitation of cyclin D1 mRNA revealed an approximate 4-fold increase at 24 hours after reperfusion. CONCLUSIONS: Aberrant expression of cell cycle-related genes may play an important role in the cell death that accompanies retinal ischemia-reperfusion injury.
Authors: Kevin C Chan; Yu Yu; Shuk Han Ng; Heather K Mak; Yolanda W Y Yip; Yolandi van der Merwe; Tianmin Ren; Jasmine S Y Yung; Sayantan Biswas; Xu Cao; Ying Chau; Christopher K S Leung Journal: Acta Biomater Date: 2019-06-06 Impact factor: 8.947
Authors: M Vidal-Sanz; M Lafuente; P Sobrado-Calvo; I Selles-Navarro; E Rodriguez; S Mayor-Torroglosa; M P Villegas-Perez Journal: Neurotox Res Date: 2000 Impact factor: 3.911