Saccon, R; (2015) Characterisation of human and mouse SOD1-ALS proteins in vivo and in vitro. Doctoral thesis , UCL (University College London).

Preview

Text Rachele Saccon PhD thesisCOMPLETE.pdf Download (12MB) | Preview

Abstract

Amyotrophic lateral sclerosis (ALS) is a fatal progressive neurodegenerative disease affecting motor neurons (MNs). It is primarily sporadic, however a proportion of cases are inherited and of these ~20 % are caused by mutations in the superoxide dismutase 1 (SOD1) gene. The work described in this thesis has focused on the characterisation of the role that the SOD1 protein plays in ALS, investigating the human and the mouse variants in vivo and in vitro. SOD1 mutations result in ALS by an unknown gain of function mechanism, although mouse models suggest that complete loss of SOD1 is also detrimental to MN function. To investigate a possible role of SOD1 loss of function in SOD1-ALS, a meta-analysis was carried out on the literature reviewing measures of SOD1 activity from patients carrying SOD1 familial ALS mutations and the phenotype of Sod1 knockout mice. The first set of experiments aimed to phenotypically characterise a novel mouse model, Sod1D83G, carrying a pathological mutation in the mouse Sod1 gene. Sod1D83G/D83G mice have no SOD1 activity, low levels of SOD1 protein, develop central MN degeneration and a distal peripheral neuropathy. Further the Sod1D83G mice were crossed with Sod1 knockout mice and mice overexpressing the human wild-type SOD1 to determine if it was possible to dissect elements of a loss of function (the peripheral axonopathy) and aspects of a gain of function (the central body degeneration). ALS mutations generally cause SOD1 to become more aggregate-prone, but it is unclear whether the human and the mouse SOD1 proteins co-aggregate in mouse models of SOD1 familial ALS. To investigate possible interactions between human and mouse SOD1 variants, recombinant proteins were produced, characterised and their spontaneous aggregation propensity was assessed in vitro. Finally a sensitised screen focused on the effect of unknown mutations on the life-span of a low copy SOD1G93A transgenic model of ALS, identified one mouse line with reduced survival, named Galahad. The phenotype of the Galahad mouse progeny was examined and a quantitative trait loci analysis was carried out to try to identify possible modifying locus/loci interacting with the SOD1 G93A mutation. The present work aims to shed light on the interaction between human and mouse SOD1 proteins and increase our understanding on the mechanism affecting central and peripheral degeneration of MNs in the context of SOD1 familial ALS mutations.

Download activity - last month

Export as CSVJSONXML

Download activity - last 12 months

Export as XMLCSVJSON

Downloads by country - last 12 months

Export as JSONCSVXML

Archive Staff Only

View Item