Histopathological evaluation of retinal damage during intraocular hypertension in rabbit: involvement of ganglion cells and nerve fiber layer.

Abstract

Current clinical tests do not detect glaucomatous signs until the onset of substantial retinal damage. Therefore animal models are required to investigate the very early histopathological alterations in glaucoma disease. We used an experimental model of intraocular hypertension to compare early changes in retinal ganglion cell (RGC) density with the thickness of the nerve fiber layer (NFL). Methylcellulose 2% was injected into the anterior chamber of 18 eyes of 18 New Zealand albino rabbits. Intraocular pressure was measured 6 h after the injection and thenceforth once a day using a Shiötz tonometer. Histopathological analysis was performed on days 4, 10, and 15 following the induction of hypertension (six eyes for each group). Sections from the upper temporal retina were stained with hematoxylin-eosin and immunohistochemically using a polyclonal antibody PGP 9.5 to identify RGC. An image analysis system was used to evaluate the RGC and the thickness of the NFL. We observed a significant increase in intraocular pressure until the end of the experiment. Histological analysis showed, after 10 days of ocular hypertension, a significant decrease in RGC density (P < 0.05) and a significant increase (P < 0.05) in glial cell density. We found a significant correlation between RGC loss and cell area at 4 days (P < 0.01; Cc = 0.86) and at 10 days (P < 0.002; Cc = 0.91) of intraocular hypertension. We did not observe a significant decrease in the NFL thickness until 10 days of intraocular hypertension. Our study confirms the size-dependent RGC loss during intraocular hypertension and shows no early decrease in NFL thickness.