Experiments involving immunostaining and cell number quantification were evaluated in an unbiased and blinded manner. Briefly, stained slides that comprised the optic nerve region were evaluated in the central retina. Both dorsal and ventral regions were counted to encompass the entire retinal region, spanning approximately ~300-400mm from the optic nerve in either direction. Regions of extreme structural damage or nearer to the ciliary marginal zone (CMZ) were excluded from counts as to not cause artificially elevated numbers. Fluorescent cell counts from two independent sections were counted and averaged from each eye. For gap counting, slides were stained with HuC/D primary antibodies and To-Pro-3 nuclei marker and regions of missing retinal ganglion cells were counted under a fluorescent microscope. Significance was calculated using a two-way ANOVA with Tukey’s post-hoc test for inter-group comparisons in GraphPad Prism 9.4.1. All data represented in graphical form is of the average ± standard error of the mean (s.e.m). The number of fish used in each experiment is described in the figure legends.