Molecular Analysis of Pax3 in the Development of Neural Crest Cells

Abstract

Neural crest cells, a transient population of cells that exists only during embryonic development, migrate from the neural tube to many different locations in the embryo where these cells are able to differentiate into a wide variety of terminal cell types, including cardiac cells, adipocytes, neurons, and melanocytes. Pax3 is one transcription factor known to play a role in the induction of neural crest development and the terminal differentiation of migrated neural crest cells. Pax3 also plays a role in skeletal muscle development, and in this lineage it has been shown to be a weak transcriptional activator and may require the binding of cofactors to increase its efficiency, but the only cofactors isolated to date are corepressors that reduce its activity. In order to study the cofactors of Pax3, we first began optimizing the culture conditions to efficiently generate neural crest cells from Pax3 mouse embryonic stem cells. Here we show that when comparing neural crest growth in an embryoid body system and a monolayer system, differentiating stem cells in a monolayer may be a better system for the generation of neural crest cells. While results are preliminary, they indicate that more experiments need to be performed to generate the optimum culture conditions. This would allow for further experiments aimed at isolating the cofactors of Pax3, which are imperative to identify in order to better understand the function of Pax3 in neural crest development and to better understand the processes and genes that govern the development of these cells.