Quantitative analysis of the effect of leflunomide on neural crest cell gene regulation during early embryonic development

The neural crest is a transient population of cells that arises at the border between the neural and non-neural ectoderm. These cells are induced, undergo an epithelial-to-mesenchymal transition, and then migrate along stereotypical pathways to form an array of derivatives such as pigment cells, cranio-facial cartilage and sensory neurons. Neural crest cells have long been studied and much about these cells and their interactions is still not fully understood. The small molecule compound leflunomide inhibits neural crest development. Leflunomide’s mode of action is to inhibit pyrimidine biosynthesis, thereby, preventing RNA transcription. Neural crest genes are actively transcribed and like many embryonic stem cells and tumour cells genes undergo an increased level of transcriptional pausing and subsequent elongation making a number of these genes sensitive to leflunomide. It was unclear at what stage of neural crest development leflunomide was acting. Here, I initially developed a quantitative approach using real-time PCR to measure gene expression in Xenopus. Secondly, using real-time PCR I have shown that neural plate border genes are not affected by leflunomide. Thirdly, the neural crest specification genes are affected and the pan neural plate marker Sox2 is not affected by leflunomide. I have confirmed by quantitative real-time PCR that the expression of genes involved in neural crest specification the proto-oncogene cMyc and cMyc responsive genes are affected. cMyc is implicated in embryonic stem cell transcriptional elongation and is well characterised to play an important role in neural crest specification.

Name dc:type.qualificationname

other

Level dc:type.qualificationlevel

masters

Grantor dc:publisher.institution

University of East Anglia

Year dc:date.issued

2014

Author dc:creator

• Ford, Christopher

Language dc:language

en