Expression of polysialylated NCAM but not L1 or N-cadherin by regenerating adult mouse optic fibers in vitro.

This study asks if there might be irreversible maturational changes in adult neurons that limit their capacity to regenerate. Retina from adult and embryonic mouse were placed in culture on laminin substrates so that regenerating adult optic fibers could be compared to growing embryonic fibers. Several cell adhesion molecules (CAMs) known to mediate the growth of embryonic neurites on astrocytes were assayed by immunocytochemistry: L1, N-cadherin, and NCAM. Thy 1.2, a potential CAM with inhibitory activity, was also examined. As in vivo, embryonic fibers were found to express both L1 and N-cadherin. In contrast, regenerating adult fibers had no detectable amounts of either of these CAMs. N-Cadherin is normally down regulated during development so its absence in adult fibers suggests it can not be reexpressed during regeneration. L1 is normally found in the proximal regions of adult optic fibers so its absence indicates it is not expressed or transported in regenerating fibers. Adult regenerating fibers expressed high levels of Thy 1.2, which was undetectable in embryonic optic fibers. Thy 1.2 is normally found in mature fibers, indicating this phenotypic feature is preserved during regeneration. Both adult and embryonic fibers showed strong reactivity for NCAM, which in vivo is normally found in embryonic and at lower levels in adult fibers. Surprisingly, both embryonic and regenerating adult fibers expressed high levels of polysialic acid, which is normally absent in adult fibers. NCAM may be one of few CAMs available to adult optic fibers for regeneration on astrocytes.

Histopathologic consequences of moderate concussion in an animal model: correlations with duration of unconsciousness.

Although duration of unconsciousness is commonly used as a prognostic index following traumatic brain injury (TBI), few controlled studies have statistically evaluated the relationship between unconsciousness and histologic pathology, particularly after moderate head injury. Using a pendulum-striker concussive device, a reproducible model of TBI in rats was developed. This model is uncomplicated by skull fractures, contusions, or experimenter-induced craniotomies. In the present study, the severity of the histopathology observed in this model of moderate closed-head injury at 48 h posttrauma is linearly related to the duration of unconsciousness (p < 0.0001). The pathology, assessed with a silver stain for neurodegeneration, is particularly striking if unconsciousness persists for 4 minutes or more. These data suggest that the initial period of unconsciousness may be a useful predictor of clinical brain histopathology associated with moderate closed-head injury, predicting either the degree of pathology and/or the rate it progresses if left untreated.

Injury induced expression of growth-associated protein-43 in adult mouse retinal ganglion cells in vitro.

In optic fibers, as in most axons of the central nervous system, the axonal growth-associated protein, GAP-43, is abundant during development but absent in adults. Since optic fibers can be induced to regenerate in culture, we examined whether this was associated with an increased expression of GAP-43 in adult mouse optic fibers that were regenerating from organotypic retinal explants on to laminin substrates. We found that simply placing adult mouse retina in culture under serum-free conditions was sufficient to induce GAP-43, which was detectable after about four to five days in vitro, coincident with the initiation of neurite outgrowth. In explants taken from animals in which the optic nerve was crushed in the orbit eight days prior to culturing, GAP-43 was observed within one day, as was neurite outgrowth. This priming effect was also seen in vivo as an increased level of GAP-43 reactivity in retinal ganglion cells and optic fibers in histological sections taken eight days after nerve crush. Reactivity in the adult fibers in culture was comparable to that observed in optic neurites growing from embryonic retinal explants and could be maintained for at least four weeks in culture. In the adult neurites, especially with longer times in culture, GAP-43 tended to be concentrated into varicosities that were often found in terminal-like arbors that formed in culture. Placing adult retina in culture under serum-free conditions in sufficient to induce re-expression of GAP-43 for an indefinite period of time. This suggests that GAP-43 expression and the propensity for growth in vivo may be repressed by a factor that is absent in vitro.