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The Use of Molecular Techniques to Investigate Genetic Targets for Enhancing CNS Neurite Outgrowth

Thomas Haynes Hutson

出版	University of London, 2010
URL	http://books.google.com.hk/books?id=rLNAtwAACAAJ&hl=&source=gbs_api

註釋

Following spinal cord injury central nervous system neurons show a very limited regenerative response, which results in their failure to successfully form functional connections with their original target. This is due in part to the reduced intrinsic growth state of CNS neurons, which is characterised by their failure to express regeneration associated genes (RAGs) and the presence of growth inhibitory molecules in CNS that form a molecular and physical barrier to regeneration. However, advancements in molecular techniques have elevated gene therapy into a promising therapeutic strategy for CNS injuries. -- The aims of this thesis were to enhance the neurite outgrowth of CNS neurons firstly by increasing their intrinsic growth state, secondly by decreasing their response to myelin inhibitors and thirdly to identify a viral vector that efficiently transduces corticospinal neurons (CSNs) that project to the corticospinal tract (CST). -- To enhance the intrinsic growth state of CNS neurons a medium-throughput electroporation assay was optimised, which allowed the investigation of potential novel regeneration associated genes to increase the neurite outgrowth of postnatal cerebellar granule neurons (CGNs). These neurons were used instead of spinal or cortical neurons due to the high purity and yield of the culture preparation and led to the discovery of a novel target; PTPN2 which when over-expressed increased neurite outgrowth on both permissive and inhibitory substrates. -- To attenuate the effect of the inhibitory environment, RNA silencing was used to knockdown the expression of Lingo-1; a protein essential for the inhibitory effect of several myelin inhibitors. Utilising an efficient and potentially therapeutically applicable integration deficient lentiviral vector delivery system, CGNs were efficiently transduced and Lingo-1 expression shown to be decreased.