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，包括周围神经系统和头部的骨头和软骨。神经嵴细胞随着神经胚层进入神经管而在神经褶皱中产生，然后迁移到胚胎外围的各种远处部位。尽管神经嵴对脊椎动物的发育至关重要，但人们还不知道这些细胞是如何形成的。本建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一个排列的cDNA文库。神经嵴特异性基因将通过杂交从神经嵴诱导和未诱导的组织中提取的标记cDNA进行鉴定。从这些细胞中，神经嵴细胞身份的调节因子将通过三个标准来识别：1)序列，2)电穿孔异位表达时诱导神经嵴的能力，以及3)表达模式。在未来的工作中，通过这种技术鉴定的基因将被功能表征，以了解产生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.