GABAa receptor regulation and trafficking in C. elegans

描述（由申请人提供）： 这一开发/探索性R21提案的目标是开发一种 研究GABAA受体运输和调节的新方向 突触将采用一种遗传模式生物方法， 秀丽隐杆线虫GABAA受体是主要的抑制剂， 神经递质受体在大脑中，并发挥重要作用， 功能协调发展的突触处GABAA受体的丰度是一个重要因素 在正常的神经系统功能中，以及在神经系统的进展中， 疾病，尤其是焦虑症。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， 运输到突触和降解。然而，迄今为止， 主要是描述性的。我们缺乏对这些过程如何发生的理解， 如何进行监管。进展因困难而放慢。 干扰哺乳动物的基因表达， 哺乳动物神经系统和哺乳动物GABAA的结构异质性 受体。一种解决办法是在一个简单的模型中采用遗传学方法 有机体C. elegans非常适合这个目的：GABAA受体， 它们在突触上的调节，在C.线虫;遗传操作 梭elegans是快速和高效的; C.线虫的神经系统 相对简单; C.线虫GABAA受体是均一的， 表征了这项建议的具体目标有两个方面：第一， 指出了沿着生物合成、运输和降解途径， 将测定突触处的对照GABAA受体丰度。实现 这一目标，这些点将受到干扰遗传，和影响的 将确定受体丰度的调节。受体测量 功能（使用电生理学），受体亚细胞定位，和 将进行受体mRNA和蛋白质水平。第二， 将确定这些潜在监管点中的每一个的重要性。这 目的将通过使用功能性GFP标记的前向遗传筛选来实现。 受体的这些初步数据将构成未来实质性研究的基础。 研究计划，以了解GABAA受体丰度在突触是如何 监管.由于细胞过程在C. 线虫和人类，这些研究的见解可以改善 基于调节GABAA受体丰度的焦虑症治疗方法 神经突触

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.