Epigenetic regulation of the temporal responsiveness of human embryonic stem cells to generate motor neurons. (360G-Wellcome-085116_Z_08_Z)

I propose to ask whether there is a temporal restriction of responsiveness of human ES derived neural stem cells to patterning signals known to be sufficient to induce a MN fate with a focus on the epigenetic regulation of such restriction. Motor neurone disease (MND) is a progressive and fatal neurodegenerative condition.There is a great need for renewable sources of human motor neurons (MNs) as an experimental resource to learn more about the aetiopathogenesis of MND and to enable the deve lopment of potential therapies. The ability to generate defined neuronal cell types from hESCs offers unique experimental opportunities to study mechanism(s) underlying neural diversity. hESC derived NSCs predictably respond to morphogenetic signals permitting the generation of region specific neurons including MNs. An unresolved question is whether responsiveness to such signals is developmentally restricted. I will generate enriched populations of MNs (using established methods). Character isation will include qPCR, FACS / IHC as well as functional assays. Our hESC neuralisation system provides an ideal platform to begin characterising the epigenetic regulation of MN specification from hESC-NSCs. Analysis of the temporal responsiveness of hESC-NSCs has several important implications including defining the optimal timeframe for in vitro generation of MNs from hESC-NSCs.