A screen for factors involved in nerve migration

描述(申请人提供)：项目计划使用显影剂 非洲爪哇三叉神经节中参与调节因子的筛选 三叉神经的迁移，希望发现新的蛋白质 和机械装置。 1.建立一种基于神经突起胶原基质生长的分析方法。 非洲爪哇三叉神经节组织块。轴突生长将受到以下因素的影响 联合培养非洲爪哇动物冠层中产生的因子 合成核糖核酸。 2.利用表达克隆技术筛选具有吸引/驱避活性的因子。 通过注射动物帽子表达的蛋白质，这些RNA池来自一种 CDNA文库将在设计用于检测可溶性或 膜结合吸引剂/驱避剂。 3.确认克隆因子(S)实际参与控制 三叉神经生长。对轴突的直接影响将通过以下方式确定 转基因表达克隆因子的体外实验 培养中的细胞。原位杂交，水泥腺体替代试验，以及 具有可诱导表达的转基因胚胎将用于建立一种 活体角色。

DESCRIPTION (provided by applicant): The project plans to use the developing Xenopus trigeminal ganglion in a screen for factors involved in regulating the migration of the trigeminal nerve, with the hope of discovering new proteins and mechanisms. 1. Develop an assay based on the growth in a collagen matrix of neurites from a Xenopus trigeminal ganglion explant. Neurite growth will be influenced by factors produced by cocultured Xenopus animal caps previously injected with synthetic RNA. 2. Screen for factors with attracting/repellent activity by expression cloning. Proteins expressed by animal caps injected with pools of RNA derived from a cDNA library will be tested in conditions designed to detect soluble or membrane-bound attractors/repellents. 3. Confirm the actual involvement of the cloned factor(s) in controlling the trigeminal nerve growth. A direct effect on the axons will be determined with in vitro assays involving the expression of the cloned factor by transfected cells in culture. In situ hybridization, a cement gland substitution assay, and transgenic embryos with inducible expression will be used to establish an in vivo role.