Postembryonic development of drosophila motoneurons

果蝇运动神经元的胚胎后发育

• 批准号: 7563978
• 负责人: RICHARD B LEVINE
• 金额: $ 16.27万
• 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/7563978
• 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; Gated Ion Channel; Growth; Health; Human; In Vitro; Ion Channel; Larva; Life; Maintenance; Measurement; Measures; Molecular; Morphology; Motor Neurons; Nature; Neurons; Pathway interactions; Pattern; 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（由申请人提供）：正常的大脑功能需要精确的树突形状和选举特征。神经元必须在初始开发过程中获得这些特性，并通过生命维持它们，除非需要功能可塑性。该项目的长期目标是了解神经活性在调节树突形状和电压门控离子通道的分布中的作用。我们已经开发了遵循遗传学模型生物体果蝇果蝇中鉴定的运动神经元在变形过程中鉴定出的运动神经元的树突状发展的技术，并证明活性水平显着影响树突状发育。神经元活性和钙依赖性转导途径可以精确地在体内和原发性细胞培养中进行操纵。具体目的是：1。确定在变质过程中如何修饰已确定的飞行运动神经元的功能属性。将采用全电池电流和电压钳技术。 2.确定哪种形式的电压门控钾和钙通道在不同阶段表达，它们在运动神经元内的分布以及通过类固醇激素调节。通道表达将通过RT-PCR进行测量，并通过强迫用四型固醇基序的通道的表达来确定分布。 3。表征树突开发的actitivition依赖性调节。通过驱动改性离子通道的表达以及通过体内和体外测量生长和功能确定的效果，将通过驱动改性离子通道的表达来增加或减少鉴定运动神经元的活性模式。 4。确定钙依赖性信号通路的作用。将使用遗传编码的钙指标，神经元离子通道的高度保守性质以及调节发育过程的分子途径确定活性依赖性钙流入的来源和性质，这表明从该模型系统中得出的基本信息将提供对人类健康的见解。与公共卫生的相关性：正常的大脑功能要求神经元发展出精确的树突结构和确定的电气特征。中风和疾病（例如癫痫）可能导致这些特性的维持不当。拟议的研究旨在表征确保这些特征适当发展和维护的机制。