Characterization of Pax3 and Sox10 transgenic Xenopus laevis embryos as tools to study neural crest development

Mansour Alkobtawi,Heather Ray,Elias H Barriga,Mauricio Moreno,Ryan Kerney,Anne-Helene Monsoro-Burq,Jean-Pierre Saint-Jeannet,Roberto Mayor

doi:10.1016/j.ydbio.2018.02.020

Abstract

The neural crest is a multipotent population of cells that originates a variety of cell types. Many animal models are used to study neural crest induction, migration and differentiation, with amphibians and birds being the most widely used systems. A major technological advance to study neural crest development in mouse, chick and zebrafish has been the generation of transgenic animals in which neural crest specific enhancers/promoters drive the expression of either fluorescent proteins for use as lineage tracers, or modified genes for use in functional studies. Unfortunately, no such transgenic animals currently exist for the amphibians Xenopus laevis and tropicalis, key model systems for studying neural crest development. Here we describe the generation and characterization of two transgenic Xenopus laevis lines, Pax3-GFP and Sox10-GFP, in which GFP is expressed in the pre-migratory and migratory neural crest, respectively. We show that Pax3-GFP could be a powerful tool to study neural crest induction, whereas Sox10-GFP could be used in the study of neural crest migration in living embryos.

Highlights

The neural crest is a multipotent population of cells that arises at the border between the developing nervous system and surface ectoderm and originates a variety of cell types, which contribute to the formation of many organs and systems (LaBonne and BronnerFraser, 1999; Huang and Saint-Jeannet, 2004; Bronner and SimõesCosta, 2016; Mayor and Theveneau, 2013)
We show that Pax3-green fluorescent protein (GFP) could be a powerful tool to study neural crest induction, whereas Sox10-GFP could be used in the study of neural crest migration in living embryos
We show that Pax3-GFP transgenic embryos express GFP in early neural crest cells and respond to neural crest inductive signals, and that Sox10-GFP transgenic embryos express GFP in migrating neural crest, both fluorescent labels which are readily visible in living embryos

Summary

Introduction

The neural crest is a multipotent population of cells that arises at the border between the developing nervous system and surface ectoderm and originates a variety of cell types, which contribute to the formation of many organs and systems (LaBonne and BronnerFraser, 1999; Huang and Saint-Jeannet, 2004; Bronner and SimõesCosta, 2016; Mayor and Theveneau, 2013). This multipotent stem cell population has fascinated biologists for more than a hundred years, as it represents an ideal system to study embryonic induction, epithelialto-mesenchymal transition, cell migration, differentiation, and evolution. We show that Pax3-GFP transgenic embryos express GFP in early neural crest cells and respond to neural crest inductive signals, and that Sox10-GFP transgenic embryos express GFP in migrating neural crest, both fluorescent labels which are readily visible in living embryos

Cloning and transgene generation

Generation of Pax3-GFP transgenic embryos

Wnt signaling response of Pax3-GFP transgene

Generation and characterization of Sox10-GFP transgenic embryos

Conclusions