Epithelial stratification and placode invagination are separable functions in early morphogenesis of the molar tooth

Abstract

Ectodermal organs, which include teeth, hair follicles, mammary ducts and glands such as sweat, mucous and sebaceous glands, are initiated in development as placodes: epithelial thickenings that invaginate and bud into underlying mesenchyme. These placodes are stratified into a basal and several suprabasal layers of cells. The mechanisms driving stratification and invagination are poorly understood. Using the molar tooth as a model for ectodermal organ morphogenesis, we show here that vertical, stratifying cell divisions are enriched in the forming placode and that stratification is cell division-dependent. Using inhibitor and gain-of-function experiments, we show that FGF signalling is necessary and sufficient for stratification but not invagination as such. We show that, instead, Shh signalling is necessary for, and promotes, invagination once suprabasal tissue is generated. Shh-dependent suprabasal cell shape suggests convergent migration and intercalation, potentially accounting for post-stratification placode invagination to bud stage. We present a model in which FGF generates suprabasal tissue by asymmetric cell division while Shh triggers cell rearrangement in this tissue to drive invagination all the way to bud formation.