WNT5A-Ca²⁺-CaN-NFAT signalling plays a permissive role during cartilage differentiation in embryonic chick digit...

During digit development, the correct balance of chondrogenic signals ensures the recruitment of undifferentiated cells into the cartilage lineage or the maintenance of cells at the undifferentiated stage. WNT/β catenin maintains the pool of progenitor cells, whereas TGFβ signalling promotes cartilage differentiation by inducing Sox9 expression. Moreover, WNT5A promotes the degradation of β catenin during mouse limb development. Although these mechanisms are well established, it is still unknown whether the signalling pathway downstream WNT5A is also involved in early chondrogenesis during digit formation. Thus, the aim of this study was to determine the role of WNT5A during the recruitment of progenitor cells during digit development. Our results showed that WNT5A activated calcium (Ca²⁺) release in the undifferentiated region during digit development. Further, the blockade of Ca²⁺ release or calcineurin (CaN) or nuclear factor of activated T-cells (NFAT) functions resulted in an inhibition of cartilage differentiation. Together, our results demonstrate that non canonical WNT5A-Ca²⁺-CaN-NFAT signalling plays a key role during embryonic digit development in vivo promoting the competence for chondrogenic signals and also acts as a permissive factor for chondrogenesis independently of cell death mechanisms.