Functional Characterization of an AMPA receptor: GluR3

描述（由申请人提供）：嗜离子性谷氨酸受体（iGluR’s）是一种配体门控离子通道，介导中枢神经系统中大多数快速兴奋性神经传递。谷氨酸神经传递功能异常可导致神经系统疾病。针对谷氨酸受体设计的药物有可能治疗许多神经系统疾病。了解谷氨酸受体的结构、功能和动态之间的关系，对于开发治疗iGluR相关疾病所需的亚型选择性药物具有重要意义。对一种特定AMPA受体亚型（GluR3）的单通道电生理研究将提供不同激动剂诱导的这些受体的功能特性，并可作为未来研究其他嗜离子性谷氨酸受体的模型。这些研究的第一个目的是表征在不同配体浓度下的完全激动剂谷氨酸和一组结构相关的部分激动剂威拉地碱化合物的不同水平的电导类。第二个目标是建立通道行为模型，并确定最能代表通道激活模式的动力学速率。单通道数据将用不同的方法进行分析，使用QuB软件，其中包括一个隐藏的马尔可夫模型方法来检测事件。底物模型和动力学速率将在完全和部分激动剂之间进行比较，以确定通道功能的相似性或差异性。将大鼠GluR3基因稳定转染到HEK 293细胞中，采用细胞贴壁法测量单通道电流。数据将获得EPC7放大器和模拟低通滤波与贝塞尔滤波器在最大10 kHz。通过了解这些通道在配体结合域结构背景下的动力学，可以设计新的药理学药物来针对特定的功能成分。公共卫生相关性：许多急性和慢性神经系统疾病涉及离子渗透性谷氨酸受体的功能。然而，有关这些受体功能的有限知识限制了新的选择性药理学药物的发展。了解谷氨酸受体的各个方面，如结构和功能，将导致更好的药物设计和更好的治疗大量患有神经系统疾病的人群。

DESCRIPTION (provided by applicant): Ionotropic glutamate receptors (iGluR's) are ligand gated ion channels that mediate most of the fast excitatory neurotransmission in the central nervous system. Aberrant function of glutamate neurotransmission can lead to neurological disorders. Drugs designed to target glutamate receptors have the potential to treat a number of neurological conditions. Understanding the relationship between the structure, function and dynamics of glutamate receptors is Important for the development of subtype selective pharmacological agents necessary to treat iGluR related disorders. Single channel electrophysiological studies completed on one specific AMPA receptor subtype (GluR3) will provide functional properties of these receptors induced by different agonists and can be used as a model for future studies on other ionotropic glutamate receptors. The first objective of these studies is to characterize the various levels of conductance classes at different ligand concentrations for a full agonist, glutamate, and a group of structurally related partial agonists, the willardiine compounds. The second objective is to model channel behavior and determine kinetic rates that best represent the mode of channel activation. Single channel data will be analyzed with different methods using QuB software that includes a hidden Markov model approach to event detection. The substrate model and kinetic rates will be compared between the full and partial agonists to determine any similarities or differences in channel function. The rat GluR3 gene has been stably transfected into HEK 293 cells and single channel currents will be measured using the cell attached method. Data will be obtained with an EPC7 amplifier and analog low pass filtered with a Bessel filter at a maximum of 10 kHz. By understanding the kinetics of these channels in the context of the structure of the ligand binding domain, new pharmacological agents can be designed to target a specific component of function. PUBLIC HEALTH RELEVANCE: Numerous acute and chronic neurological conditions involve the function of ion permeable glutamate receptors. However, limited knowledge pertaining to the function of these receptors restricts development of new selective pharmacological agents. Understanding various aspects of glutamate receptors, such as structure and function, will lead to better drug design and better treatment for the large population that suffer from neurological conditions.