TWIST1: a subtle modulator of neural differentiation and neural tube formation

Abstract

The central nervous system is formed from epiblast precursor cells through Neurulation. Neural induction can be studied in its main aspects in vitro. However, the process is poorly understood, especially in regard to when and how a cell becomes specified, and then committed, to be a neural cell. It is, on the other hand, well established that neural formation requires absence or, inhibition of the BMP signalling both in vivo and in vitro. ID1 is a direct target of BMP signalling with major influence on in vitro neural differentiation. A cDNA library screen, looking for transcription factors negatively regulated by ID1, reported TWIST1, along with only two other proteins. Twist1 expression is upregulated during in vitro neural differentiation. Furthermore, targeted deletion of Twist1 has dramatic consequences on anterior neural development. Twist1 knock-out mice fail to form the closed neural tube in the prospective brain, followed by exencephaly and, early embryonic death. In this thesis I investigate the influence on in vitro neural differentiation of a TWIST1 constitutively active form, insensitive to ID1 inhibition. I report that this transcriptionally active TWIST1 accelerates neural differentiation, in vitro and, biases it, towards dorsal phenotypes. I provide, for the first time, evidence for Twist1 expression in the neural tissue, observed weakly in a restricted domain, temporally and spatially, in the dorsal part of the neural tube. I propose a new model for TWIST1 influence at this level. I also investigate how TWIST1 actions depend on levels of expression and dimer choice. I found that, TWIST1 can exert its neural modulating actions only at low levels, as high levels divert a cell fate towards non-neural lineages.