The Extreme Anterior Domain Is an Essential Craniofacial Organizer Acting through ...

• The Kinin-Kallikrein pathway regulates craniofacial development
• Nitric oxide regulates mouth and neural crest development
• Bradykinin stimulates nitric oxide production in the embryo
• The extreme anterior domain (EAD) attracts first arch neural crest using Cpn

The extreme anterior domain (EAD) is a conserved embryonic region that includes the presumptive mouth. We show that the Kinin-Kallikrein pathway is active in the EAD and necessary for craniofacial development in Xenopus and zebrafish. The mouth failed to form and neural crest (NC) development and migration was abnormal after loss of function (LOF) in the pathway genes kng, encoding Bradykinin (xBdk), carboxypeptidase-N ( cpn), which cleaves Bradykinin, and neuronal nitric oxide synthase ( nNOS). Consistent with a role for nitric oxide (NO) in face formation, endogenous NO levels declined after LOF in pathway genes, but these were restored and a normal face formed after medial implantation of xBdk-beads into LOF embryos. Facial transplants demonstrated that Cpn function from within the EAD is necessary for the migration of first arch cranial NC into the face and for promoting mouth opening. The study identifies the EAD as an essential craniofacial organizer acting through Kinin-Kallikrein signaling.