Abstract

The neural crest (NC) is a major contributor to the vertebrate craniofacial skeleton, detailed in model organisms through embryological and genetic approaches, most notably in chick and mouse. Despite many similarities between these rather distant species, there are also distinct differences in the contribution of the NC, particularly to the calvariae of the skull. Lack of information about other vertebrate groups precludes an understanding of the evolutionary significance of these differences. Study of zebrafish craniofacial development has contributed substantially to understanding of cartilage and bone formation in teleosts, but there is currently little information on NC contribution to the zebrafish skeleton. Here, we employ a two–transgene system based on Cre recombinase to genetically label NC in the zebrafish. We demonstrate NC contribution to cells in the cranial ganglia and peripheral nervous system known to be NC–derived, as well as to a subset of myocardial cells. The indelible labeling also enables us to determine NC contribution to late–forming bones, including the calvariae. We confirm suspected NC origin of cartilage and bones of the viscerocranium, including cartilages such as the hyosymplectic and its replacement bones (hymandibula and symplectic) and membranous bones such as the opercle. The cleithrum develops at the border of NC and mesoderm, and as an ancestral component of the pectoral girdle was predicted to be a hybrid bone composed of both NC and mesoderm tissues. However, we find no evidence of a NC contribution to the cleithrum. Similarly, in the vault of the skull, the parietal bones and the caudal portion of the frontal bones show no evidence of NC contribution. We also determine a NC origin for caudal fin lepidotrichia; the presumption is that these are derived from trunk NC, demonstrating that these cells have the ability to form bone during normal vertebrate development.

Highlights

  • The evolution of vertebrates is concomitant with the evolution of the multi–potent neural crest (NC), which contributes to much of the vertebrate craniofacial skeleton

  • We have developed an approach to indelibly label the NC cells and their descendants, using a two– transgene system based on Cre recombinase

  • We used a recently described enhancer associated with the mouse Sox10 gene, which has been analyzed through transgenesis in mouse and zebrafish [18,24]

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Summary

Introduction

The evolution of vertebrates is concomitant with the evolution of the multi–potent neural crest (NC), which contributes to much of the vertebrate craniofacial skeleton. Data in the frog suggests that the entire vault of the skull contains NC-derived cells [5], unlike the situation in mouse or that supported by some of the data in chick. In neither mouse nor chick is there any evidence that trunk NC cells gives rise to cartilage or bone during normal development. Smith and colleagues demonstrated migration of trunk NC into the caudal fin mesenchyme [8]. The authors speculated these cells might contribute to the bony lepidotrichia, but lacked the lineage data to demonstrate that

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