Gene expression profiling of GABA neurons to reveal synaptic remodeling genes

GABA 神经元的基因表达谱揭示突触重塑基因

• 批准号: 8039974
• 负责人: Sarah Petersen
• 金额: $ 1.57万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2011-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8039974
• 关键词: Address; Adopted; Biological Assay; Biological Models; Bypass; Caenorhabditis elegans; Candidate Disease Gene; Cells; Cholinergic Receptors; Cues; Data Set; Defect; Development; Dorsal; Employee Strikes; Erinaceidae; Event; Family; Foundations; Gene Expression; Gene Expression Profiling; Genes; Genetic; Goals; Indium; Lead; Learning; Locomotion; Memory; Methods; Molecular; Morphology; Motor; Motor Neurons; Muscle; Nematoda; Nervous system structure; Neuronal Plasticity; Neurons; Nuclear Hormone Receptors; Pathway interactions; Phosphotransferases; Process; Proteins; RNA Interference; Role; Signal Transduction; Specificity; Synapses; Synaptic plasticity; Testing; Therapeutic; Transcript; chicken ovalbumin upstream promoter-transcription factor; cholinergic neuron; design; gamma-Aminobutyric Acid; gene function; hatching; human disease; in vivo; member; molecular polarity; mutant; novel; postsynaptic; presynaptic; prevent; programs; protein function; research study; synaptogenesis; transmission process

DESCRIPTION (provided by applicant): Neurons adopt polarized morphologies to direct information flow in the nervous system. The assembly of distinct presynaptic and postsynaptic regions is necessary for the transmission of signals from one neuron to the next. These domains may be remodeled by developmental events that alter both axonal and dendritic compartments. The capacity of neurons to remodel polarity is evolutionarily conserved but the mechanisms that govern this process are largely unknown. The GABAergic motor neurons in the nematode C. elegans display a striking example of developmentally regulated synaptic remodeling. Dorsal D (DD) motor neurons initially innervate ventral muscles but switch polarity after hatching to synapse with dorsal muscle. Ventral D (VD) motor neurons that arise during this period are prevented from remodeling by UNC-55, a member of the conserved family of COUP transcription factors. The goal of this project is to exploit this model system to identify molecular factors that govern motor neuron remodeling. In Aim 1, I will test the hypothesis that a novel mechanism, independent of known synaptogenic proteins SYD-1 and SAD-1, drives DD motor neuron remodeling and that UNC-55 prevents VD motor neurons from adopting this pathway. To identify the potentially novel determinants of DD rewiring, I have used a powerful cell-specific microarray profiling method to detect ~200 transcripts that regulated by UNC-55 in vivo. In experiments described in Aim 2, I will use specific RNAi-dependent assays to test these candidate genes for roles in DD synaptic remodeling. A pilot screen of selected genes in this data set revealed an independent role for Hedgehog-related genes in synaptic assembly. Aim 3 is designed to define the mechanism of Hedgehog-related protein function in GABA motor neuron synaptogenesis. In this study, I expect to uncover conserved elements in the mechanism of synaptic remodeling. Thus, these results could lead to significant advances in our understanding of synaptic plasticity and thereby provide a foundation for developing therapeutic approaches for human diseases that disrupt synaptic assembly.

描述（由申请人提供）：神经元采用两极分化的形态来导致神经系统中的信息流。不同的突触前和突触后区域的组装对于从一个神经元传输到下一个信号是必需的。这些领域可以通过改变轴突和树突隔室的发展事件来重塑。神经元重塑极性的能力在进化上是保守的，但是控制这一过程的机制在很大程度上是未知的。线虫C.秀丽隐杆线虫中的GABA能运动神经元显示了发育调节的突触重塑的一个引人注目的例子。背侧D（DD）运动神经元最初支配腹侧肌肉，但在孵化后与背肌突触后转换极性。在此期间出现的腹侧D（VD）运动神经元被UNC-55（保守的政变转录因子家族的成员）防止重塑。该项目的目的是利用该模型系统来识别控制运动神经元重塑的分子因素。在AIM 1中，我将检验以下假设：一种与已知的突触源蛋白SYD-1和SAD-1无关的新型机制驱动DD运动神经元重塑，而UNC-55则可以防止VD运动神经元采用该途径。为了识别DD重新布线的潜在新型决定因素，我使用了强大的细胞特异性微阵列分析方法来检测约200个由UNC-55在体内调节的转录本。在AIM 2中描述的实验中，我将使用特定的RNAi依赖性测定测试这些候选基因在DD突触重塑中的作用。该数据集中选定基因的试验屏幕揭示了刺猬相关基因在突触组件中的独立作用。 AIM 3旨在定义GABA运动神经元突触发生中与刺猬相关蛋白功能的机理。在这项研究中，我希望在突触重塑的机理中发现保守的元素。因此，这些结果可能会导致我们对突触可塑性的理解的重大进展，从而为开发破坏突触组装的人类疾病的治疗方法提供了基础。