An Optic Nerve Crush Injury Murine Model to Study Retinal Ganglion Cell Survival

Abstract

Injury to the optic nerve can lead to axonal degeneration, followed by a gradual death of retinal ganglion cells (RGCs), which results in irreversible vision loss. Examples of such diseases in human include traumatic optic neuropathy and optic nerve degeneration in glaucoma. It is characterized by typical changes in the optic nerve head, progressive optic nerve degeneration, and loss of retinal ganglion cells, if uncontrolled, leading to vision loss and blindness.The optic nerve crush (ONC) injury mouse model is an important experimental disease model for traumatic optic neuropathy, glaucoma, etc. In this model, the crush injury to the optic nerve leads to gradual retinal ganglion cells apoptosis. This disease model can be used to study the general processes and mechanisms of neuronal death and survival, which is essential for the development of therapeutic measures. In addition, pharmacological and molecular approaches can be used in this model to identify and test potential therapeutic reagents to treat different types of optic neuropathy.Here, we provide a step by step demonstration of (I) Baseline retrograde labeling of retinal ganglion cells (RGCs) at day 1, (II) Optic nerve crush injury at day 4, (III) Harvest the retinae and analyze RGC survival at day 11, and (IV) Representative result. ProtocolAll equipments and reagents used are sterile. All animal experiments were approved by the Animal Care and Use Committee (ACUC) at the NEI/NIH (animal study protocol NEI-570), and were performed according to the NIH guidelines and regulations. Baseline Retrograde Labeling of Retinal Ganglion Cells (RGCs) At Day 1The purpose of this procedure is to label the retinal ganglion cells retrogradely by injecting a neural tracer dye into the superior colliculus of the mice three days before the optic nerve crush injury. The dye will be retrogradely taken up by the retinal ganglion cells and provides a marker for the living RGCs (Figure 1). This approach yields reproducible labeling of viable RGCs with little variation 1-5 . Optic Nerve Crush Injury At Day 4In this procedure, we will apply a crush injury to the optic nerve to cause a primary damage to the axons (Figure 2), which will lead to a gradual death of the retinal ganglion cells.

Bifidobacterium promotes retinal ganglion cell survival by regulating the balance of retinal glial cells

“…To explore the effects of gut microbiota on RGCs, mice were fed different antibiotics (ampicillin (1 mg/mL), colistin (1 mg/mL), or vancomycin (0.5 mg/mL)) for 21 days. On day 7 of antibiotic intervention, the mice were subjected to ONC, a classic model of optic nerve injury that induces RGC death 22 . On day 21 of antibiotic intervention, the mice were sacrificed, and their retinas were isolated (Figure 1A,B).…”

“…On day 7 of antibiotic intervention, the mice were subjected to ONC, a classic model of optic nerve injury that induces RGC death. 22 On day 21 of antibiotic intervention, the mice were sacrificed, and their retinas were isolated (Figure 1A,B).…”

Comparative Proteomic Study of Retinal Ganglion Cells Undergoing Various Types of Cellular Stressors

“…The murine optic nerve crush injury model is the most commonly used model of RGC injury that mimics molecular events occurring upon traumatic optic neuropathy, glaucoma, etc. (Tang et al, 2011) Moreover, the prevalence of optic neuropathy in diabetics could increase with diabetes duration (Hua et al, 2019) In this model, the crush injury to the optic nerve leads to retinal ganglion cell apoptosis. This disease model can be used to study the general processes and mechanisms of neuronal death and survival, which is essential for the development of therapeutic measures.…”

DNA Damage and Repair in Eye Diseases

“…Furthermore, persistent DNA damage response might be responsible for aging and neurodegenerative diseases such as Alzheimer’s disease and amyotrophic lateral sclerosis [ 93 ]. Clinically, optic nerve crush injury models are similar to glaucoma to the extent that retinal ganglion cell (RGC) death is a main pathologic phenomenon [ 94 ]. DNA damage responses can be attenuated through the inhibition of Mre11 in the MRN complex or ataxia telangiectasia mutated (ATM) kinase.…”