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  • Title: Expression of cell cycle-related genes in dying cells in retinal ischemic injury.
    Author: Kuroiwa S, Katai N, Shibuki H, Kurokawa T, Umihira J, Nikaido T, Kametani K, Yoshimura N.
    Journal: Invest Ophthalmol Vis Sci; 1998 Mar; 39(3):610-7. PubMed ID: 9501873.
    Abstract:
    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.
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