Molecular identification of distinct neurogenic and melanogenic neural crest sublineages

Abstract

Clonal and lineage analyses have demonstrated that although some neural crest cells have the ability to generate multiple cell types and display self-renewal ability, other crest cells generate a single or limited repertoire of cell types. However, it is not yet clear when, and in what order, crest cells become specified to adopt a particular fate. We report that the receptor tyrosine kinases TrkC and C-Kit are expressed by distinct neural crest subpopulations in vitro. We then analyzed the lineages of individual receptor-expressing crest cells and found that TrkC-expressing cells that have just emerged from the neural tube give rise to clones containing neurons or glial cells, or both, but never produce melanocytes. A short time later, TrkC-expressing cells only generate pure neuronal clones. By contrast, from their earliest appearance in neural tube outgrowths, C-Kit-expressing cells invariably give rise to clones containing only melanocytes. Our results directly demonstrate that distinct neurogenic and melanogenic sublineages diverge before or soon after crest cells emerge from the neural tube, that fate-restricted precursors are present in nascent neural crest populations and that these sublineages can be distinguished by their cell type-specific expression of receptor tyrosine kinases

Neural crest cells retain their capability for multipotential differentiation even after lineage‐restricted stages

“…7). In contrast to our studies, an in vitro culture study using quail embryo NC cells showed that the Kitþ NC cells invariably give rise to clones containing only M (Luo et al, 2003). Additionally, an in vivo immunohistochemistry study on mouse embryos showed that the migrating Kitþ NC cells give rise to only M (Wilson et al, 2004).…”

“…NC-like cells were isolated as Kit-expressing cells by flow cytometry, and they differentiated into N, G, and M, suggesting that these Kit-expressing cells have multipotency. However, in vivo Kit-expressing NCderived cells are reported to migrate dorsolaterally in the skin and differentiate only into M (Henion and Weston, 1997;Luo et al, 2003;Wilson et al, 2004). To further investigate the commitment potential of these early NClike cells, we took advantage of the Sox10 transcription factor as a wellcharacterized NC marker.…”

“…The Sox10-positive cells that were generated in a very low number in the presence of Noggin were sorted and re-cultured on ST2 stromal cells. Although a few colonies were formed, no N, G, and M were generated from these Sox10-expressing cells (data not shown (Luo et al, 2003;Wilson et al, 2004). To examine the discrepancy between the differentiation potential of ES cell-derived Sox10þ NC-like cells tested in vitro and that of their embryonic counterpart, we generated mice from the Sox10-IRESVenus ES cells.…”

“…Clonal analysis of NC cells shows that almost half of the initial NC cell population is a lineage-restricted population, which generates clones of only a single cell type (Henion and Weston, 1997), or is a population comprising bipotent and unipotent precursors (Lahav et al, 1998). In particular, a subpopulation of early NC cells that express the receptor tyrosine kinase Kit exclusively express M-lineage markers in rodents (Wilson et al, 2004), and avian Kit-expressing NC cells invariably give rise to clones containing only melanocytes (Henion and Weston, 1997;Luo et al, 2003). Furthermore, none of the ventrally migrating NC cells express Kit (Wilson et al, 2004), and they do not differentiate into M even under conditions that favor melanocyte development (Reedy et al, 1998).…”

The proliferating field of neural crest stem cells

“…Hence, it will be interesting to see whether eNCSCs and pigment cell precursors segregate from a truly multipotent neural crest stem cell. Such a scenario would be consistent with the identification of segregated melanocyte progenitors in mouse dorsal neural tube (Wilson et al, 2004) and early migrating NCCs from avian neural tube explants (Henion and Weston, 1997;Luo et al, 2003).…”