DEVELOPMENT OF NEURONAL FUNCTION AND FORM

大分子结构和细胞机械是 在基因的指导下构建的。适当的收藏品 突变体应该使我们能够剖析结构或机制 参与神经元的发育和功能。建议数 利用果蝇突变体研究基本神经元 神经功能与形态的机制及其相互关系 在开发过程中。 一种新的果蝇幼虫中枢神经系统细胞培养体系 允许直接的形态观察和生理 记录发育中的神经元。我们将继续努力 鉴定培养的幼虫神经元以获得基线信息 来研究这个系统中的神经生物学问题。神经元 具有不同形态模式的人可以在培养中被分类 源自各种已定义的中枢神经系统区域。染色方式可用 抗幼虫中枢神经系统抗体和单抗的研制 将用于细化分类。区别和 控制离子通道功能的基因表达重叠 将在不同的细胞类型中进行检查。特别是它的影响 几个K+通道突变，其中差异表达 建议用膜片钳方法进行研究。 为了跟进我们的初步调查结果， 不同性别间神经突起生长模式的超兴奋性 用K+进一步分析培养神经元的类型 渠道变种人。SHI/TS基因突变对细胞膜的影响 循环，导致生长锥体活动和轴突停止 超出29摄氏度的热脉冲将应用于 含有少量SHI/TS感觉神经元的遗传嵌合体 评估发育迟缓和胎膜的影响 轴突导航和终末分枝的再循环停滞 和竞争。 ShI/ts基因产物将通过凝胶电泳法鉴定 研究其生化特性和亚细胞定位。至 从这个蛋白质延伸到其他功能相关的成分 在控制膜回收的分子网络中，我们将 分离缺陷抑制或增强SHI/TS的新突变体 表型。

Macromolecular structures and cellular machineries are constructed under direction of genes. A proper collection of mutants should enable us to dissect the structures or mechanisms involved in neuronal development and function. The proposed study employs Drosophila mutants to study basic neuronal mechanisms and relationships between neuronal function and form during development. A recently developed cell culture system of Drosophila larval CNS allows direct morphological observation and physiological recording of developing neurons. We will continue our effort to characterize cultured larval neurons to gain baseline information for studies of neurobiological problems in this system. Neurons with distinct morphological patterns can be classified in cultures derived from various defined CNS regions. Staining by available antibodies and monoclonal antibodies raised against larval CNS will be used to refine the classification. The distinction and overlap of expression of genes controlling ion channel function will be examined among cell types. Particularly, the effects of several K+ channel mutations, of which differential expression has been suggested, will be studied by patch clamp methods. To follow up on our preliminary findings, the influence of hyperexcitability on neurite outgrowth pattern among different types of cultured neurons will be further analyzed by using the K+ channel mutants. Mutations of shi/ts gene affect membrane recycling, resulting in arrest of growth cone activity and neurite extension above 29 degrees C. Heat pulses will be applied to genetic mosaics containing a few shi/ts sensory neurons to evaluate the effects of developmental delay and membrane recycling arrest on axonal navigation and terminal arborization and competition. The shi/ts gene product will be identified by gel electrophoresis to study its biochemical properties and subcellular localization. To extend from this protein to other functionally related components in the molecular network controlling membrane recycling, we will isolate new mutants whose defects suppress or enhance the shi/ts phenotype.