Abstract
BackgroundNeural crest cells (NCCs) are embryonic, multipotent stem cells. Their long-range and precision-guided migration is one of their most striking characteristics. We previously reported that P0-Cre/CAG-CAT-lacZ double-transgenic mice showed significant lacZ expression in tissues derived from NCCs.ResultsIn this study, by embedding a P0-Cre/CAG-CAT-EGFP embryo at E9.5 in collagen gel inside a culture glass slide, we were able to keep the embryo developing ex vivo for more than 24 hours; this development was with enough NCC fluorescent signal intensity to enable single-cell resolution analysis, with the accompanying NCC migration potential intact and with the appropriate NCC response to the extracellular signal maintained. By implantation of beads with absorbed platelet-derived growth factor-AA (PDGF-AA), we demonstrated that PDGF-AA acts as an NCC-attractant in embryos.We also performed assays with NCCs isolated from P0-Cre/CAG-CAT-EGFP embryos on culture plates. The neuromediator 5-hydroxytryptamine (5-HT) has been known to regulate NCC migration. We newly demonstrated that dopamine, in addition to 5-HT, stimulated NCC migration in vitro. Two NCC populations, with different axial levels of origins, showed unique distribution patterns regarding migration velocity and different dose-response patterns to both 5-HT and dopamine.ConclusionsAlthough avian species predominated over the other species in the NCC study, our novel system should enable us to use mice to assay many different aspects of NCCs in embryos or on culture plates, such as migration, division, differentiation, and apoptosis.
Highlights
Neural crest cells (NCCs) are embryonic, multipotent stem cells
Another example of interspecies differences is seen in the pathways of cranial neural crest cells (CNCCs) migration in mammals, which are not nearly as well delineated as they are in birds [29]
All the results of this study demonstrated the usefulness of the P0-Cre/CAG-CAT-EGFP reporter system for various NCC analyses
Summary
Neural crest cells (NCCs) are embryonic, multipotent stem cells. Their long-range and precisionguided migration is one of their most striking characteristics. In mammals, NCCs begin to emigrate from the tip or ‘crest’ of the still-open neural folds [27], whereas in birds NCCs arise only after the neural tube closure occurs [28]. Another example of interspecies differences is seen in the pathways of CNCC migration in mammals, which are not nearly as well delineated as they are in birds [29]. Fish or frog embryos exhibit markedly different patterns of CNCC emigration from mammals or birds
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