GABAa receptor regulation and trafficking in C. elegans

线虫中 GABAa 受体的调节和运输

• 批准号: 6418012
• 负责人: Bruce A Bamber
• 金额: $ 14.45万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-12-10 至 2003-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6418012
• 关键词: Caenorhabditis elegans; GABA receptor; alternatives to animals in research; biochemical evolution; chimeric proteins; electrophysiology; endocytosis; genetic manipulation; genetic models; genetic regulation; genetic screening; genetic transcription; green fluorescent proteins; intracellular transport; muscimol; neuroregulation; neurotransmitter agonist; northern blottings; protein biosynthesis; protein degradation; protein localization; protein structure function; protein transport; receptor expression; synapses

DESCRIPTION (provided by applicant): The objective of this developmental/exploratory R21 proposal is to develop a new direction for studying GABAA receptor trafficking and regulation at synapses. A genetic model organism approach will be taken, using the nematode Caenorhabditis elegans. GABAA receptors are the major inhibitory neurotransmitter receptors in the brain, and play important roles in most brain functions. The abundance of GABAA receptors at synapses is an important factor in normal nervous system function, and in the progression of nervous system disease, particularly anxiety disorders. Studies in mammalian systems suggest that receptor abundance is regulated by controlling receptor biosynthesis, trafficking to synapses, and degradation. However, results to date have been largely descriptive. We lack an understanding of how these processes occur, and how they may be regulated. Progress has been slowed by the difficulty of perturbing mammalian gene expression, the anatomical complexity of the mammalian nervous system, and the structural heterogeneity of mammalian GABAA receptors. One solution is to take a genetic approach in a simple model organism. C. elegans is well suited for this purpose: GABAA receptors, and their regulation at synapses, are conserved in C. elegans; genetic manipulation of C. elegans is rapid and efficient; the C. elegans nervous system is relatively simple; and the C. elegans GABAA receptor is uniform and well characterized. The specific aims of this proposal are two-fold: First, the points along the biosynthetic, trafficking, and degradation pathways that control GABAA receptor abundance at synapses will be determined. To achieve this aim, these points will be perturbed genetically, and the effects on the regulation of receptor abundance will be determined. Measurements of receptor function (using electrophysiology), receptor subcellular localization, and receptor mRNA and protein levels will be performed. Second, the genes that are important at each of these potential regulatory points will be determined. This aim will be achieved by forward genetic screening using a functional GFP-tagged receptor. These initial data will form the basis of a substantial future research program to understand how GABAA receptor abundance at synapses is regulated. Because cellular processes are generally well-conserved between C. elegans and humans, insights from these studies could lead to improved treatments for anxiety disorders based on modulating GABAA receptor abundance at synapses.

描述（由申请人提供）： 该开发/探索性 R21 提案的目标是开发一个 研究GABAA受体运输和调控的新方向 突触。将采用线虫遗传模型生物方法 秀丽隐杆线虫。 GABAA受体是主要的抑制性受体 大脑中的神经递质受体，在大多数大脑中发挥重要作用 功能。突触处 GABAA 受体的丰度是一个重要因素 正常神经系统功能以及神经系统的进展 疾病，特别是焦虑症。对哺乳动物系统的研究表明 受体丰度是通过控制受体生物合成来调节的， 运输到突触和退化。然而，迄今为止的结果是 很大程度上是描述性的。我们对这些过程如何发生缺乏了解，并且 如何对它们进行监管。进展因困难而减慢 扰乱哺乳动物基因表达，解剖学的复杂性 哺乳动物神经系统和哺乳动物GABAA的结构异质性 受体。一种解决方案是在简单模型中采用遗传方法 生物。线虫非常适合这个目的：GABAA 受体，以及 它们对突触的调节在秀丽隐杆线虫中是保守的；基因操纵 线虫的检测快速高效；线虫神经系统是 相对简单；线虫 GABAA 受体均匀且良好 特点。该提案的具体目标有两个：第一， 生物合成、运输和降解途径上的点 控制突触处的 GABAA 受体丰度将被确定。达到 为了实现这一目标，这些点将受到遗传干扰，并对 受体丰度的调节将被确定。受体的测量 功能（使用电生理学）、受体亚细胞定位，以及 将进行受体 mRNA 和蛋白质水平检测。其次，基因 将确定这些潜在监管点的重要性。这 目标将通过使用功能性 GFP 标记的正向遗传筛选来实现 受体。这些初始数据将构成未来实质性的基础 了解突触 GABAA 受体丰度的研究计划 受监管。因为细胞过程在 C. 线虫和人类，这些研究的见解可能会导致改进 基于调节 GABAA 受体丰度的焦虑症治疗 在突触处。