Dr Alison Twelvetrees
The coordination of microtubule-dependent motors during axonal transport.
Neurodegenerative disease in general
Alzheimer | Amyotroph | Dementia | Huntington | Motor Neuron
My aim is to understand how a direct interaction between the kinesin and dynein motors affects the activity and motility of each, at both the single molecule and cellular level, to discover how this regulates the delivery of axonal cargoes. Initially, I will use biochemical techniques to characterise the interaction between kinesin-1 and dynein and then use this information to target mutations in the kinesin-1 protein. Multiple parameters will then be used to measure the motility of single molecules of fluorescently-labelled dynein and kinesin motors in vitro via a range of microscopy techniques. Following on from these single molecule methods, I will develop embryonic stem (ES) cells that express exogenous dynein with kinesin-1 binding domain mutations in the absence of endogenous dynein (which will be deleted through Cre-mediated recombination). Movements of dynein motors and axonal cargoes will then be analysed in motor neurons differentiated from mutant ES cells. If these experimen ts are successful, transgenic mice will be generated from the ES cells with the endogenous dynein subunit deleted only in specific subsets of neurons (e.g. sensory and motor neurons) so that axonal transport can be imaged in vivo in the sciatic nerve.