Myelin injury plays a role in many devastating conditions, including multiple sclerosis, and neuromyelitis optica. Previous work in understanding myelin injury has utilised a combination of high-resolution static imaging modalities, such as electron microscopy, and ensemble-averaged imaging, such as immunohistochemistry. Within the work presented in this thesis the behaviour of myelin basic protein was addressed, during various stages of differentiation and injury.
In order to explore this several reporters for myelin basic protein were designed and encoded within Semliki forest constructs in order to induce protein transduction in oligodendroglial cells. Reporters utilised several fluorescent tags including Dendra2, mCherry, and GFP. These viral vectors were characterised to determine the expression kinetics and toxicity within cells.
Following the production of the MBP reporters, a complement injury modality was optimised to induce myelin/oligodendrocyte specific injury with little injury within axons/neurones. This myelin injury was utilised for live imaging of myelin using the MBP reporters produced previously.
The reporters produced were utilised to explore the change in particle dynamics following myelin injury, using single molecule TIRF imaging. Responses to injury were compared over various stages of oligodendrocyte differentiation, where it was found that MBP dynamics following injury varied most when mature oligodendrocytes were injured
