Optic Nerve Crush Injury Model

Male and female mice at age seven to eight weeks were randomly assigned to the treatment groups. ONC injury was performed as described previously (Park et al., 2008 (link); Patel et al., 2017 (link)). The mice were anesthetized using an isoflurane mixture and one drop of 0.5% proparacaine hydrochloride was added to the eye. A small incision was made in the superior posterior area of the conjunctiva to expose the optic nerve. ONC was performed ∼1 mm behind the globe carefully avoiding blood vessels for 5 s with extra-fine self-closing forceps. The ONC eyes were intravitreally injected with recombinant Wnt5a (20 and 50 ng, in 2 μl; R&D Systems Inc) or the equivalent volume of sterile saline, using a 33-gauge Hamilton needle (Hamilton Company). Erythromycin ointment was topically applied to treat the injected eye and analgesia was provided with buprenorphine SR (0.5 mg/ml). Animals were excluded from the study if they had excessive bleeding or swelling at any time after the procedure.