Date of Award

8-2012

Degree Type

Thesis

Degree Name

Cell and Molecular Biology (M.S.)

Department

Cell and Molecular Biology

Abstract

Floor plate cells of the mesencephalon have the ability to become more floor plate cells or differentiate into dopamine containing neurons. Maturation of these neurons can be marked by the stage of differentiation. When dopamine neurons of the substantia nigra pars compacta are no longer able to communicate with the corpus striatum due to a marked loss in the neurotransmitter dopamine, movement disorders such as those present in Parkinson’s disease occur. The bHLH transcription factor Nato3 is known to be necessary for normal dopamine neuron formation, however, our lab was interested in its sufficiency to drive a dopaminergic lineage. The aim of this study was to begin to understand the molecular mechanisms responsible for dopamine neurogenesis in order to understand therapeutic means that may help combat Parkinson’s disease when these dopamine neurons are lost. Overexpression of Nato3 as well as cofactor combinations (Otx2, Foxa2 and Lmx1a) was performed using in ovo electroporation and a fast screening method of qPCR was implemented in order to screen multiple genes that may be involved in the production of dopamine (Nurr1, Pitx3 and TH). Unfortunately, due to variability and inconsistencies of the electroporation efficiency and the low magnitude of change in marker expression, it is inconclusive as to whether or not Nato3 is sufficient to drive dopamine neurogenesis. Multiple approaches to improve the variability in the methods are addressed.

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