PURPOSE. This study investigated the role of sterile alpha/Armadillo/Toll-Interleukin receptor homology domain 1 protein (SARM1) in Wallerian-like degeneration of retinal ganglion cells (RGCs) and their axons after inducing excitotoxicity. METHODS.To induce excitotoxicity, kainic acid (KA) was injected into the vitreous humor of B6.Cg-Tg(Thy1-YFP)HJrs/J mice. Control mice received PBS. At 24, 48, and 72 hours after injection, degeneration of RGCs and their axons in the retina was determined by fundus imaging, and axonal degeneration in the optic nerves was determined by fluorescence microscopy. SARM1 protein levels were determined by Western blot analysis and SARM1 tissue localization was determined by immunohistochemistry. Causal role of SARM1 in KAmediated degeneration of RGCs and their axons was determined by treating the eyes with KA along with Sarm1 silencer siRNA. RESULTS.Fundus imaging and microscopic analysis indicated that KA promoted Wallerian-like degeneration of RGCs and axons in KA-treated eyes, but not in PBS-treated eyes. Quantitative analysis indicated a significant increase in degeneration of RGCs and their axons in KA-treated injected eyes, but not in PBS-treated eyes. Compared with low levels of SARM1 protein in retinal protein extracts, retinal cross sections, and optic nerve from PBS-treated eyes, SARM1 protein levels were increased in KA-treated eyes. Finally, treatment of eyes with KA along with a Sarm1 silencer siRNA attenuated KA-mediated degeneration of RGCs and their axons significantly. CONCLUSIONS.Results presented in this study, for the first time, show that KA-mediated upregulation of SARM1 protein promotes Wallerian-like degeneration of RGCs and their axons.