NEURONAL DEVELOPMENT

这份提案概述了一系列旨在测试和完善 我们对神经系统发育的机制的理解 被精确地刻画了图案。这样的精确度对于正常的 大脑的功能；产生这种模式的手段是 对我们理解正常的胎儿发育和对 开发从疾病和损伤中恢复的疗法 神经系统的创伤。研究人员的实验能力 然而，分析脊椎动物的大脑发育受到 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，这项提案概述了 正在进行的一系列实验，以检验模式的发展 果蝇翅膀中的周围神经系统 黑猩猩。在果蝇的翅膀上，PNS的个体元素 以一种非常刻板的方式从一张上皮片上长出来。 系统的相对简单性和一些可用的 分子和基因工具使其非常有利于实验 分析。我们将使用遗传、分子和 免疫组织学技术分析这一疾病的分子基础 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，并开始搜索 对构图过程至关重要的新基因。 这个项目的具体目标是：L)进一步发展方法 利用分子标记技术分析发育中翅的致死突变 携带染色体和FRT-FLP系统，并开发新的方法 诱导所选基因在不连续的部分中过度表达 2)描述Achaete-Scute复合体(AS-C)、螺旋-C的详细事件 环螺旋(HLH)与感受器神经源性基因表达 用免疫组织化学方法观察翼状叶片的前体发育 技术；3)测试“抑制”hlh基因在控制中的作用 利用嵌合体分析和三七总皂苷特异性研究正常感受器的发育 细胞标志物；4)实验分析细胞的性质--细胞 负责精炼的相互作用内的“原神经”区域 机翼叶片，使用遗传和外科技术；5)检查 Saggy，一种丝氨酸苏氨酸激酶，在建立位置信息中的作用 在叶片上，通过探测已知和新的基因 转录被毛茸茸的突变改变；以及6)分离和 利用增强子陷阱果蝇分析新的位置调控基因 台词。

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.