Source Paper
Lens Injury Stimulates Axon Regeneration in the Mature Rat Optic Nerve
Steven Leon, Yuqin Yin, Jennifer Nguyen, Nina Irwin, Larry I. Benowitz
Journal of Neuroscience • 2000
View Abstract
In mature mammals, retinal ganglion cells (RGCs) are unable to regenerate their axons after optic nerve injury, and they soon undergo apoptotic cell death. However, a small puncture wound to the lens enhances RGC survival and enables these cells to regenerate their axons into the normally inhibitory environment of the optic nerve. Even when the optic nerve is intact, lens injury stimulates macrophage infiltration into the eye, Müller cell activation, and increased GAP-43 expression in ganglion cells across the entire retina. In contrast, axotomy, either alone or combined with intraocular injections that do not infringe on the lens, causes only a minimal change in GAP-43 expression in RGCs and a minimal activation of the other cell types. Combining nerve injury with lens puncture leads to an eightfold increase in RGC survival and a 100-fold increase in the number of axons regenerating beyond the crush site. Macrophage activation appears to play a key role, because intraocular injections of Zymosan, a yeast cell wall preparation, stimulated monocytes in the absence of lens injury and induced RGCs to regenerate their axons into the distal optic nerve.
Lens Injury and Optic Nerve Crush
Objective: Assessment of retinal ganglion cell survival and axon regeneration following combined lens puncture wound and optic nerve crush injury in mature rats
Gather these items before starting the experiment. Check off items as you prepare.
Materials1
Not specified • Not applicable • Not mentioned • Not mentioned
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Protocol Steps
Lens Puncture Wound
Create a small puncture wound to the lens
Note: Even when optic nerve is intact, lens injury stimulates macrophage infiltration into the eye
View evidence from paper
“a small puncture wound to the lens enhances RGC survival and enables these cells to regenerate their axons”
Optic Nerve Crush Injury
Perform optic nerve crush injury
Note: Axotomy alone causes only minimal change in GAP-43 expression in RGCs
View evidence from paper
“axotomy, either alone or combined with intraocular injections that do not infringe on the lens, causes only a minimal change in GAP-43 expression”
Combined Lens Injury and Nerve Crush
Combine nerve injury with lens puncture wound
Note: Combined injury produces synergistic effects on RGC survival and axon regeneration
View evidence from paper
“Combining nerve injury with lens puncture leads to an eightfold increase in RGC survival and a 100-fold increase in the number of axons regenerating beyond the crush site”
Intraocular Zymosan Injection
Inject Zymosan intraocularly to stimulate monocytes and induce RGC axon regeneration without lens injury
Note: Zymosan stimulates monocytes in absence of lens injury and induces RGCs to regenerate axons into distal optic nerve
View evidence from paper
“intraocular injections of Zymosan, a yeast cell wall preparation, stimulated monocytes in the absence of lens injury and induced RGCs to regenerate their axons into the distal optic nerve”