Distinct ossification trade-offs illuminate the shoulder girdle reconfiguration at the water-to-land transition.

The mechanisms of the pectoral girdle transformation at the origin of terrestrial locomotion in vertebrates remain an outstanding problem. The loss of intramembranous bones and the enlargement of endochondral bones resulted in the disarticulation of the pectoral girdle from the skull and the formation of the neck during the fish-to-tetrapod transition. Despite the functional implications of this skeletal shift in the emergence of terrestrial vertebrates, the underlying genetic-developmental alterations have remained enigmatic. Here, we show that in zebrafish pectoral girdle mesodermal cells expressing gli3, a transcription factor gene in the Hedgehog signaling pathway, differentiate into both intramembranous and endochondral bones. Intriguingly, Gli and Hedgehog compound knockout fish exhibited an unexpected combination of actinopterygian fish and stem-tetrapod pectoral girdle characteristics. These ontogenetic and anatomical data suggest that a trade-off between the two distinct ossification pathways is a deeply embedded developmental program in bony fishes and that tuning of this trade-off can generate novel pectoral girdle akin to those of stem-tetrapods at the dawn of vertebrate terrestrialization.