Evaluation of Midbrain Dopaminergic Neuron Specification in Rat ES Cell Derived Neurons
Summary
Parkinson's disease (PD) is a chronic, neurodegenerative disorder, which arises as a result of the progressive loss of midbrain dopaminergic (mDA) neurons in the substantia nigra. The severity of the symptoms negatively influences the patients’ quality of life and comprises an economic burden to the government. Although many treatments have been developed over time, these therapies only reduce symptoms and often have side effects. Since PD results from the loss of a specific cell type, it lends itself to cell replacement therapy. A very promising source for cell replacement therapy are embryonic stem (ES) cells. The use of rat ES cells in this perspective, offers some advantages compared to the utilization of mouse, human or primate ES cells. Rat ES cells have been reported to differentiate into neurons by means of adjusting the level of MEK/ERK inhibition. With regard to PD, it would be interesting to know if these neurons have differentiated toward a midbrain or even mDA neuronal fate. During this project, the RT-PCR conditions for markers for midbrain specification and mature mDA neurons were optimized and neuronal samples, previously generated by modulation of the level of MEK/ERK inhibition, were evaluated for midbrain marker expression by RT-PCR analysis. Indeed markers of mDA neuronal specification were identified in rat ES cell derived neurons. This result provides useful knowledge for future experiments.