A screen for factors involved in nerve migration

筛选参与神经迁移的因素

• 批准号: 6798287
• 负责人: ALIN VONICA
• 金额: $ 5.58万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-01-11 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6798287
• 关键词: Xenopus; cell growth regulation; cell migration; collagen; complementary DNA; developmental neurobiology; embryo /fetus; expression cloning; ganglions; gene expression; genetic techniques; genetically modified animals; in situ hybridization; molecular genetics; nerve /myelin protein; neurogenesis; neurogenetics; nucleic acid sequence; protein structure function; technology /technique development; tissue /cell culture; transfection; trigeminal nerve

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.

描述（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