Constitutively active Notch1 converts cranial neural crest-derived frontonasal mesenchyme to perivascular cells $\textit{in vivo}$

Abstract

Perivascular/mural cells originate from either the mesoderm or the cranial neural crest. Regardless of their origin, Notch signalling is necessary for their formation. Furthermore, in both chicken and mouse, constitutive Notch1 activation (via expression of the Notch1 intracellular domain) is sufficient $\textit{in vivo}$ to convert trunk mesoderm-derived somite cells to perivascular cells, at the expense of skeletal muscle. In experiments originally designed to investigate the effect of premature Notch1 activation on the development of neural crest-derived olfactory ensheathing glial cells (OECs), we used $\textit{in ovo}$ electroporation to insert a tetracycline-inducible $\textit{NotchΔE}$ construct (encoding a constitutively active mutant of mouse Notch1) into the genome of chicken cranial neural crest cell precursors, and activated $\textit{NotchΔE}$ expression by doxycycline injection at embryonic day 4. $\textit{NotchΔE}$-targeted cells formed perivascular cells within the frontonasal mesenchyme, and expressed a perivascular marker on the olfactory nerve. Hence, constitutively activating Notch1 is sufficient $\textit{in vivo}$ to drive not only somite cells, but also neural crest-derived frontonasal mesenchyme and perhaps developing OECs, to a perivascular cell fate. These results also highlight the plasticity of neural crest-derived mesenchyme and glia.