Dhasmana, Devesh J; (1999) The role of the Schwann cell in the induction of elongative central axonal growth. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

The factors underlying the failure of axon regeneration in the CNS are thought to comprise of both the lack of supportive factors as well as the presence of inhibitory ones. Transplantation work has shown that the PNS is able to provide some of the necessary criteria and with it an increased capacity to regenerate. Studies have further shown that the crucial ingredient to such peripheral grafts is the presence of Schwann cells (SCs), the major glial cell of the PNS. I have used an extrusion transplantation system, recently developed in this laboratory, to study the effects of a SC column placed into the origin of the septo-hippocampal cholinergic projection. These SC columns integrate with host glia with minimal tissue damage, form a tight and ordered column with aligned cellular processes, and are able to recruit modest numbers of axons. Immunostaining with a cholinergic axon marker suggests that these axons arise from the septal nuclei. Given the limited availability and yield of primary SCs that current preparation protocols offer, I have engineered neonatal SC lines by transfecting the SV40 large T antigen into a population of primary neonatal rat SCs. Characterisation of these cell lines, with the use of immunocytochemistry, Western blotting and RT-PCR, shows that they retain the immunophenotype of primary SCs in vitro, although in vivo studies have posed more difficult with the lack of a suitable marker. In addition, I have set up a retroviral transfection system with the use of a bicistronic vector containing the Green Fluorescent Protein. This would provide a means of unique and efficient labelling prior to transplantation, and moreover offer the potential for further transfections of an additional gene of interest within the same vector.

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