Sodium nitroprusside (SNP), an NO donor, was studied for its effects on apoptosis in rat retinal neurons. TUNEL-positive cells were observed in the outer nuclear layer (ONL), but not in the inner retina after SNP treatment. Inner retinal neurons died by necrosis. No photoreceptor cells were found in the ONL after seven days. Immunoblotting confirmed that neurnal NO synthase expression increased up to 5 days (approximately 170% of control levels), and then declined by 7 days, suggesting that NO induces apoptosis in the ONL, and that inner retinal neurons die by necrosis due to glutamate from damaged photoreceptors.

We investigated the expression and cellular localization of growth-associated protein (GAP)-43 in the rat retina following ischemia induced by transiently increased intraocular pressure. In the normal retina, GAP-43 immunoreactivity was restricted to profiles in the inner plexiform layer. Following ischemia and reperfusion, immunoreactivity appeared in ganglion cells. The cell density of labeled ganglion cells peaked three days post-lesion and then decreased at seven days. Quantitative evaluation by immunoblotting confirmed that GAP-43 expression increased at three days (to 190% of control levels) and then slightly decreased at seven days. Our findings suggest that some ganglion cells have the potential to regenerate through the up-regulation of GAP-43 in the ischemic rat retina.

Using immunoblot analysis and immunocytochemistry, we investigated expression and cellular localization of endothelial nitric oxide synthase (eNOS) and proliferating cell nuclear antigen (PCNA) in the l-arginine treated ischemic rat retina. In parallel, we tested whether the blood-retinal barrier was intact by immunocytochemistry using an antiserum against IgG. In the l-arginine-treated ischemic retina, the magnitude of the increased eNOS was higher, and PCNA was expressed in endothelial cells as well as in neurons in the inner retina during the whole experimental period. Finally, IgG leakage was not detectable in the l-arginine-treated ischemic retina. Our results clearly suggest that the increased NO production by eNOS may be essential for the survival of endothelial cells in the rat retina following transient ischemia.