MOLECULAR CHARACTERIZATION OF NEURAL CREST INDUCTION

神经嵴细胞是脊椎动物胚胎中多种谱系的祖细胞，包括外周神经系统和头部的骨和软骨。当神经外胚层滚入神经管时，神经嵴细胞出现在神经褶皱中，然后迁移到胚胎周围的各种远距离部位。尽管神经嵴对脊椎动物的发育至关重要，但这些细胞是如何形成的还不清楚。这个提议的目标是定义产生神经嵴细胞的分子事件。将从神经嵴发育不同阶段的胚胎池中创建阵列cDNA文库。阵列上的神经嵴特异性基因将通过杂交减去的标记cDNA来鉴定，所述标记cDNA由神经嵴诱导与未诱导的组织制成。从该库中，神经嵴细胞身份的调节剂将通过三个标准鉴定：1）序列，2）当通过电穿孔异位表达时诱导神经嵴的能力，和3）表达模式。在未来的工作中，通过这种技术鉴定的基因将在功能上进行表征，以了解引起神经嵴细胞的级联事件。

Neural crest cells are progenitors of diverse and numerous lineages in the vertebrate embryo, including the peripheral nervous system and the bones and cartilage of the head. Neural crest cells arise in the neural folds as the neurectoderm rolls into a neural tube, then migrate to a variety of distant sites in the periphery of the embryo. Despite the fundamental importance of the neural crest for vertebrate development, it is not known how these cells form. The goal of this proposal is to define the molecular events that generate neural crest cells. An arrayed cDNA library will be created from a pool of embryos at different stages of neural crest development. Neural crest-specific genes on the array will be identified by hybridizing subtracted, labeled cDNA made from tissues that are induced versus uninduced for neural crest. From this pool, regulators of neural crest cell identity will be identified by three criteria: l) sequence, 2) ability to induce neural crest when ectopically expressed by electroporation, and 3) expression pattern. In future work, genes identified by this technique will be functionally characterized in order to understand the cascade of events that gives rise to neural crest cells.