Postembryonic development of drosophila motoneurons

果蝇运动神经元的胚胎后发育

• 批准号: 7188242
• 负责人: RICHARD B LEVINE
• 金额: $ 16.29万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7188242
• 关键词: Adult; Animal Model; Automobile Driving; Axon; Biological Metamorphosis; Biological Models; Brain; Calcium; Calcium Channel; Cells; Characteristics; Complex; Dendrites; Development; Developmental Process; Disease; Drosophila genus; Drosophila melanogaster; Ensure; Epilepsy; GYPA gene; Gated Ion Channel; Growth; Health; Human; In Vitro; Ion Channel; Larva; Life; Maintenance; Measurement; Measures; Molecular; Morphology; Motor Neurons; Nature; Neurons; Pathway interactions; Pattern; Polymerase Chain Reaction; Potassium; Potassium Channel; Primary Cell Cultures; Property; Public Health; Regulation; Research; Role; Shapes; Signal Pathway; Source; Staging; Stroke; Structure; Techniques; Testing; Transgenic Organisms; Voltage-Clamp Technics; calcium indicator; in vivo; insight; research study; response; steroid hormone; voltage

DESCRIPTION (provided by applicant): Normal brain function requires precise dendritic shape and electical characteristics. Neurons must acquire these properties during initial development and maintain them through life, except under circumstances where functional plasticity is required. The long-term objectives of this project are to understand the role of neuronal activity in regulating dendritic shape and distribution of voltage-gated ion channels. We have developed techniques for following the dendritic development of identified motoneurons during metamorphosis in the genetically-tractable model organism, Drosophila melanogaster, and demonstrated that activity levels influence dendritic development significantly. Neuronal activity and calcium-dependent transduction pathways can be manipulated precisely, both in vivo and in primary cell culture. Specific aims are: 1. To determine how the functional attributes of identified flight motoneurons are modified during metamorposis. Whole-cell current and voltage-clamp techniques will be employed. 2.To determine which forms of voltage-gated potassium and calcium channels are expressed at different stages, their distributions within the motoneuron and their regulation by steroid hormone. Channel expression will be measured by RT- PCR, and distribution determined by forcing the expression of channels tagged with a tetrad-cystiene motif. 3. Characterize the acitivity-dependent regulation of dendritic development. The activity patterns of identified motoneurons will be increased or decreased by driving the expression of modified ion channels and the effect determined with both in vivo and in vitro measurements of growth and function. 4. Determine the role of calcium-dependent signalling pathways. The source and nature of activity-dependent calcium influx will be determined using genetically-encoded calcium indicators, the highly conserved nature of neuronal ion channels and the molecular pathways that regulate developmental processes suggests that basic information derived from this model system will provide insights into human health. Relevance to public health: Normal brain function requires that neurons develop a precise dendritic structure and a defined array of electrical characteristics. Stroke and diseases, such as epilepsy, can result in improper maintenance of these properties. The proposed research seeks to characterize mechanisms that ensure proper development and maintenance of these characteristics.

描述（申请人提供）：正常的脑功能需要精确的树突形状和电特性。除非在需要功能可塑性的情况下，神经元必须在最初的发育过程中获得这些特性，并在一生中保持它们。该项目的长期目标是了解神经元活动在调节树突形状和电压门控离子通道分布中的作用。我们已经开发了一种技术，用于跟踪遗传易学模式生物黑腹果蝇（Drosophila melanogaster）变态过程中已识别的运动神经元的树突发育，并证明活动水平显著影响树突发育。在体内和原代细胞培养中，神经元活动和钙依赖的转导途径都可以被精确地操纵。具体目标是：1.；确定已识别的飞行运动神经元的功能属性在变态过程中是如何改变的。将采用全细胞电流和电压箝位技术。2.确定电压门控钾和钙通道在不同阶段的表达形式、在运动神经元内的分布及其受类固醇激素的调节。通道表达将通过RT- PCR来测量，并通过强制表达带有四体-胱氨酸基序标记的通道来确定分布。3. 描述树突发育的活性依赖调节。已识别的运动神经元的活动模式将通过驱动修饰离子通道的表达而增加或减少，并且通过体内和体外的生长和功能测量来确定效果。4. 确定钙依赖性信号通路的作用。活性依赖性钙内流的来源和性质将使用遗传编码的钙指标来确定，神经元离子通道的高度保守性质和调节发育过程的分子途径表明，从该模型系统中获得的基本信息将为人类健康提供见解。与公众健康相关：正常的脑功能需要神经元发育出精确的树突结构和明确的电特性阵列。中风和疾病，如癫痫，可导致这些特性的维护不当。拟议的研究旨在确定确保适当发展和维持这些特征的机制。