Three-dimensional image analysis of migration patterns of mouse neural crest cells

我们用三维图像分析技术研究了小鼠神经脊细胞的迁移模式，并用单抗4E9R对神经脊细胞进行了鉴定。在躯干，有三个硬化区，沿前后轴有不同的神经脊细胞迁移路径，这些区域的大小在时间上以系统的方式发生变化。这些数据表明，每个硬骨节内神经脊细胞迁移的区域差异及其时间变化产生了小鼠主干神经脊细胞的迁移模式。硫酸软骨素蛋白多糖在巩膜小体中的分布与神经脊细胞的迁移方式有关。此外，在用硫酸蛋白多糖生物合成抑制剂、氯酸钠和β-D-木糖苷处理的胚胎中，它们的迁移模式发生了改变。我们认为，硬化体中硫酸软骨素蛋白多糖分布的系统时空变化是形成小鼠干神经脊细胞迁移模式的关键。Tenascin在神经脊细胞迁移中的作用一直存在争议。Tenascin基因敲除小鼠胚胎中这些细胞的迁移模式与野生型CREST细胞的迁移模式相似。这支持了Tenascin不是主干神经脊细胞节段性生长的直接原因的观点。在头区，中脑神经脊细胞迁移到特定部位，即中脑与后脑交界处的间质内。

We have examined migration patterns of mouse neural crest cells by three-dimensional image analysis using the monoclonal antibody 4E9R to identify neural crest cells. In the trunk, three sclerotomal regions having different migratory pathways of neural crest cells were identified along the anteroposterior axis and the size of these regions temporally changed with a systematic mode. The data suggest that the regional differences of neural crest cell migration within each sclerotome and their temporal changes generate migration patterns of trunk neural crest cells in the mouse. The distribution of chondroitin sulfate proteoglycans in the scletotome was correlated with migration patterns of neural crest cells. Furthermore, their migration patterns were altered in embryos treated with the inhibitors of sulfated proteoglycan biosynthesis, sodium chlorate and beta-D-xyloside. We conclude that systematic spatiotemporally changes of the distribution of chondroitin sulfate proteoglycans in the sclerotome are a key requisite for the formation of migration patterns of mouse trunk neural crest cells. The role of tenascin in neural crest cell migration has been controversial. Migration patterns of these cells in tenascin-knockout mouse embryos were comparable with those of wild-type crest cells. This support the notion that tenascin is not directly responsible for the segmented outgrowth of trunk neural crest cells. In the cephalic region, mesencephalic neural crest cells migrated in the specific portion, that is in the mesenchyme proximal to the boundary between the midbrain and the hindbrain.