NEURONAL DEVELOPMENT

该提案概述了一系列旨在测试和完善 我们对发育中的神经系统 都是精确的图案这种精确度对于正常的 大脑的功能;这种模式产生的方式是 这对我们理解正常的胎儿发育， 开发用于从疾病和损伤引起的恢复的疗法， 神经系统的创伤研究人员通过实验 然而，分析脊椎动物的大脑发育受到限制， 脊椎动物神经系统的复杂性和制造困难 适当的实验操作。因此，本提案概述了 一系列正在进行的实验， 在果蝇翅膀的周围神经系统中， 黑腹菌在果蝇的翅膀中，PNS的单个元件 以一种非常刻板的方式从单一的上皮层产生。 该系统相对简单，并提供了一些 分子和遗传工具使其非常有利于实验 分析。我们将结合基因、分子和 免疫组织学技术来分析这种疾病的分子基础。 刻板印象 我们的分析包括新技术的发展， 测试一些已知基因的作用，并开始寻找 新的基因对图案形成过程至关重要。 该项目的具体目标是： 利用标记分析发育中翅膀的致命突变， 携带染色体和FRT-FLP系统，并开发新的方法， 诱导所选基因在细胞膜的离散部分中过表达， 翼; 2）描述详细的事件Achaete-scute复杂（AS-C），螺旋- 环螺旋（HLH）和感器过程中的神经基因表达 前体发育的翅膀叶片，使用免疫组织化学 3）测试“抑制性”HLH基因在控制 通过使用镶嵌分析和PNS特异性 细胞标记物; 4）实验分析细胞-细胞的性质 相互作用负责细化内的“原神经”区域的 翼片，使用遗传和外科技术; 5）检查的作用， 一种丝氨酸-苏氨酸激酶，在建立位置信息中的作用 在翼片中，通过探测已知的和新的基因， 转录被粗毛突变改变;和6）分离和 分析新的位置调控基因，使用增强子陷阱果蝇 线

This proposal outlines a series of experiments designed to test and refine our understanding of the mechanisms by which the developing nervous system is precisely patterned. Such precision is essential to the normal functioning of the brain; the means by which such patterning arises is essential to both our understanding of normal fetal development, and for developing therapies for the recovery from disease and injury-induced trauma to the nervous system. The ability of researchers to experimentally analyze brain development in vertebrates, however, is limited by the complexity of the vertebrate nervous system and difficulties making appropriate experimental manipulations. This proposal therefore outlines an ongoing series of experiments which examine the patterned development of the peripheral nervous system in the wing of the fruitfly, Drosophila melanogaster. In the fruitfly wing, the individual elements of the PNS arise in an extremely stereotyped fashion from a single epithelial sheet. The relative simplicity of the system and the availability of a number of molecular and genetic tools make it very favorable for experimental analyses. We will use a combination of genetic, molecular, and immunohistological techniques to analyze the molecular basis of this stereotypy. Our analysis includes the development of novel techniques, testing the roles of a number of known genes, and initiating a search for novel genes critical to the patterning process. The specific aims of this project are to: l) Further develop methods for the analysis of lethal mutations in the developing wing using marker- carrying chromosomes and the FRT-FLP system, and develop novel methods for inducing the overexpression of chosen genes in discrete portions of the wing; 2) Describe detailed events of Achaete-scute Complex (AS-C), helix- loop-helix (HLH), and neurogenic gene expression during sensillum precursor development in the wing blade, using immunohistological techniques; 3) Test the role of "inhibitory" HLH genes in controlling normal sensillum development by using mosaic analysis and PNS-specific cell markers; 4) Experimentally analyze the nature of the cell-cell interactions responsible for refinement within "proneural" regions of the wing blade, using genetic and surgical techniques; 5) Examine the role of shaggy, a serine-threonine kinase, in establishing positional information in the wing blade, by probing for known and novel genes whose transcription is altered by the shaggy mutation; and 6) Isolate and analyze novel positionally regulated genes, using enhancer trap Drosophila lines.