Optic Nerve Crush in Mice

Mice of 2–6 months of age were used for controlled optic nerve crush (CONC) after anesthetized with 100 mg/kg ketamine and 10 mg/kg xylazine. The optic nerve was surgically exposed and crushed just behind the globe for five seconds with self-closing forceps (Roboz RS-5027, Gaithersburg, MD) [8 (link), 18 (link)]. Ophthalmic ointment containing neomycin, polymyxin b sulfates, and dexamethasone was applied to both eyes following the procedure (Sandoz, Princeton, NJ). Control eyes included those that underwent sham surgery (optic nerve exposed but not crushed) and those eyes that were not manipulated except receiving antibiotic ointment (naïve eyes) as optic nerve crush is known to lead to microglial alterations in the contralateral eye [26 (link), 27 (link)]. Eyes were then harvested for RNA extraction and immunohistochemistry.

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Syc-Mazurek S.B., Yang H.S., Marola O.J., Howell G.R, & Libby R.T. (2022). Transcriptional control of retinal ganglion cell death after axonal injury. Cell Death & Disease, 13(3), 244.

Publication 2022

dexamethasone

Antibiotic Dexamethasone Eyes Forceps Immunohistochemistry Ketamine Mice Microglial Neomycin Nerve crush Ointment Optic nerve Polymyxin b sulfates Seconds Surgery Xylazine

Corresponding Organization :

Other organizations : University of Rochester Medical Center, Jackson Laboratory

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Variable analysis

independent variables

Controlled optic nerve crush (CONC)

dependent variables

RNA extraction
Immunohistochemistry

control variables

Mice age (2-6 months)
Anesthesia (100 mg/kg ketamine and 10 mg/kg xylazine)
Ophthalmic ointment (neomycin, polymyxin b sulfates, and dexamethasone)

controls

Positive control: Sham surgery (optic nerve exposed but not crushed)
Negative control: Naïve eyes (not manipulated except receiving antibiotic ointment)

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