We investigated the effect of brain-derived neurotrophic factor (BDNF) on retinal ganglion cell (RGC) survival after intraocular pressure (IOP) elevation at various time intervals. In adult Wistar rats, RGCs were labeled with 5% Fluorogold. Animals with 1.8-2.5-fold increase in IOP after cauterization of three episcleral vessels, were divided into three BDNF groups and three vehicle control groups, each receiving one, two or three injections. The RGC survival percentage on RGCs of the first, second and third injections were 93.9% (n = 7), 91.3% (n = 7), 82.7% (n = 5), respectively in BDNF groups; 91.6% (n = 6), 84.1% (n = 6) and 73.5% (n = 5), respectively in vehicle controls. The second and third injections of BDNF showed statistically significant survival effects. These findings demonstrated that BDNF has partial neuroprotection on RGCs in whole retina and enhances RGC survival in moderately chronic hypertensive eyes.