Regeneration in the rat optic nerve after cold injury

In order to examine nerve regeneration under conditions in which the basal laminae of the glial limiting membranes (GLM) and blood vessels were preserved intact, the intraorbital segment of adult rat optic nerve was frozen locally. During the next 3 months, degenerative and regenerative changes in axons and glial cells were observed by light and electron microscopy. On the day after treatment, all the myelinated and unmyelinated axons in the central zone of the lesion were damaged. The astrocyte endfeet of the GLM as well as the blood vessels were extensively disrupted, while their basal laminae were preserved apparently intact as a continuous sheet. Three days after treatment, regenerating axons appeared in the central zone of the lesion. They contained various numbers of clear and dense-cored vesicles as well as some smooth endoplasmic reticulum. The regenerating axons gradually increased in number, especially beneath the pial and perivascular surfaces of the lesion, where an abundance of regenerating axons was found 3 months after treatment. A few of these axons were abnormally remyelinated by oligodendrocytes. In addition to this axonal regeneration through the intraoptic nerve compartment, fine regenerating axons were seen growing out through GLM into the pial connective tissue 3 weeks after treatment. Astrocyte endfeet of the GLM became irregular in contour, protruding in a fern-leaf fashion into the pial connective tissue. Fine naked axons grew out through these protrusions and subsequently increased in number, vigorously growing in large bundles both proximally and distally along blood vessels in the pial connective tissue. Bundles of regenerating axons extended as much as 1.5 mm from the site of the lesion 3 months after surgery. These bundles were covered by thin processes of pial or arachnoidal non-neuronal cells, and the regenerating axons remained unmyelinated. The above findings indicate that under well-nourished conditions, adult mammalian optic nerve exhibits considerable regenerative ability.