A screen for factors involved in nerve migration

描述（由申请人提供）：该项目计划使用开发 非洲爪蟾三叉神经节在筛选参与调节 三叉神经的迁移，希望发现新的蛋白质 和机制。 1.开发一种基于神经突在胶原基质中生长的测定方法， 爪蟾三叉神经节外植体。神经突生长将受到以下因素的影响 共培养的非洲爪蟾动物帽产生的因子， 合成RNA。 2.通过表达克隆筛选具有引诱/驱避活性的因子。 用来自一种哺乳动物的RNA池注射动物帽所表达的蛋白质 cDNA文库将在设计用于检测可溶性或 膜结合吸引剂/排斥剂。 3.确认克隆因子实际参与控制 三叉神经生长对轴突的直接影响将由 涉及通过转染细胞表达克隆因子的体外测定 细胞培养。原位杂交，骨水泥腺替代试验，和 具有诱导表达的转基因胚胎将被用于建立一个在 体内作用

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.