TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK
Dopamine neurons are functionally important neurons, yet the genetic mechanisms that program their correct development are poorly understood. In the parent grant to this administrative supplement, we have proposed to isolate and characterize genes involved in dopamine neuron development, using the nematode C.elegans as a model system. These genes have been identified through forward genetic screens in which mutants with defects in dopamine neuron developmment have been isolated. The key challenge is to identify the nature of the gene(s) mutated in these strains. In this administrative supplement, we propose to further develop whole genome shotgun sequencing (WGS) as a cost-effective, fast and reliable approach to shortcut traditional mapping approaches. We have requested funds to upgrade an exististing Illumina Genome Analyzer II machine, which will allow us to more expediently undertake full genome sequencing of mutant strains, defective in dopamine neuron development. Furthermore, we have requested funds to undertake genome sequencing of a collection of dopamine (-) mutant animals, retrieved through additional forward genetic screens.
We have done the upgrade as requested and we have performed whole genome sequencing on a number of mutants. This work identified a novel homeobox factor involved in dopamine neuron specification. We expect that together with other factors that we previously identified, we can generate dopamine neuron types in an in vitro system. This bears strong promise for cellular replacement therapies for Parkinson's disease.