Use this URL to cite or link to this record in EThOS:
Title: A proteomic approach to studying oligodendrocyte signalling
Author: Chan, C. W.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 2007
Availability of Full Text:
Full text unavailable from EThOS.
Please contact the current institution’s library for further details.
In this study, my aim was to investigate oligodendrocyte signalling using a non-candidate, unbiased, proteomic approach. First, in collaboration with my industrial sponsors Merck Sharpe and Dohme, the potential of using 2-dimensional differential in gel electrophoresis (2D DIGE) to study oligodendrocyte signalling was evaluated. Although differentially expressed proteins between the 2 proteomes under analysis could be identified, it was concluded that 2D DIGE would be an unsuitable method to study oligodendrocyte biology. An alternative approach was therefore pursued in which a specific subset of the proteome was analysed. Previously, α6β1 integrin has been shown to be involved in oligodendrocyte survival and morphological differentiation. A protocol was developed whereby protein complexes co-immunoprecipitated with α6β1 integrin could be identified by mass spectroscopy analysis. Multiple proteins were identified including β1 integrin, contactin, plectin and heterogeneous ribonuclear protein K (hnRNP K). Characterisation and function of contactin in oligodendrocyte α6β1 integrin signalling was then investigated. Contactin expression throughout oligodendrocyte in vitro differentiation was confirmed by both Western blotting and Immunocytochemistry. The interaction of contactin and α6β1 was also verified by traditional co-immunoprecipitation. siRNA knockdown of contactin had no effect on in vitro oligodendrocyte morphological differentiation. However, contactin siRNA knockdown did block oligodendrocyte enhanced survival of the α6β1 integrin substrate, laminin-2, in response to platelet derived growth factor (PDGF). Therefore, by taking a proteomic approach, I have identified a novel interaction between α6β1 integrin and contactin, and a potential role of contactin in α6β1 integrin mediated survival signalling.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available