SoxE genes are multifunctional transcriptional regulators that play key roles in specification and differentiation of neural crest. Three members (Sox8, Sox9, Sox10) are expressed in the neural crest and are thought to modulate the expression and activity of each other. In addition to regulating the expression of other early neural crest marker genes, SoxE genes are required for development of cartilage. Here we investigated the role of SoxE genes in development of the neural crest-derived branchial skeleton in the sea lamprey. Using a morpholino knockdown approach, we show that all three SoxE genes described in lamprey are required for branchial basket development. Our results suggest that SoxE1 and SoxE2 are required for specification of the chondrogenic neural crest. SoxE3 plays a morphogenetic role in patterning of the branchial basket and may be required for the development of mucocartilage, a tissue unique to larval lampreys. While the lamprey branchial basket develops primarily from an elastin-like major extracellular matrix protein that is specific to lampreys, fibrillar collagen is also expressed in developing branchial cartilage and may be regulated by the lamprey SoxE genes. Our data suggest that the regulation of Type II collagen by Sox9 might have been co-opted by the neural crest in development of the branchial skeleton following the divergence of agnathan and gnathostome vertebrates. Finally, our results also have implications for understanding the independent evolution of duplicated SoxE genes among agnathan and gnathostome vertebrates.

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