Axon regeneration in organotypic slice cultures from the mammalian auditory system is topographic and functional

In vitro models have frequently been employed to investigate the specificity of the formation of axonal projections during both development and regeneration. Such studies demonstrated pathway, target, and laminar specificity, yet they did not tackle the problem of topography. Here, we addressed the issue of regeneration of spatial specificity at the topographic level by lesioning a precisely organized projection from the auditory system of neonatal rats in organotypic slice culture and by analyzing regeneration capacity. Lesioning had no effect on the survival of axotomized neurons or the structure of the auditory nuclei. Anterograde and retrograde biocytin tracing demonstrated that the projection regenerated topographically at the supracellular level. Whole-cell patch-clamp recordings revealed that the regenerated projection was functional. Topographic regeneration was not impaired by blocking spike activity with tetrodotoxin or glycinergic transmission with strychnine. However, if lesioning was performed after the slices had been incubated for 1 week, regeneration capacity was lost despite good survival of neurons. The loss of the regeneration capacity in vitro occurs at a developmental stage that corresponds to the age when the capacity for axonal reorganization is lost in vivo. We conclude that the developmental processes occurring in vivo and in vitro are comparable in this system, which is why we think that essential aspects of the loss of regeneration capacity may be addressed with our culture model in the future.