HUMAN GABA RECEPTORS

人类 GABA 受体

• 批准号: 6187886
• 负责人: ROBERT A DAVIDOFF
• 金额: $ 24.55万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-15 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6187886
• 关键词: GABA receptor; Xenopus oocyte; afferent nerve; electrophysiology; embryo /fetus tissue /cell culture; gamma aminobutyrate; human fetus tissue; human genetic material tag; laboratory rat; neurons; neuropharmacology; neurotransmitter antagonist; picrotoxin; spinal ganglion; voltage /patch clamp

DESCRIPTION: This proposal aims to investigate the electrophysiological/ pharmacological characteristics and subunit composition of GABA receptors located on human primary afferent neurons. The proposed research stems from intriguing results of recent investigations which establish that cultured human primary afferent neurons express GABA receptors with unique, and hitherto undescribed, pharmacological properties. An association with both a large transplant center and an internationally recognized spinal cord trauma unit has provided the applicants with access to human neurons. Three separate laboratories with expertise in: (1) neuronal cell culturing; (2) electrophysiology and pharmacology of GABA receptors; and (3) molecular biology of neurotransmitter receptors will participate in these studies. The overall intent is to clarify the properties of these novel human GABA receptors by using a combination of sophisticated electrophysiological techniques, pharmacological probes, and molecular biological methods so as to define the channel/molecular properties and to determine the GABA receptor subunits in human sensory neurons. The proposed investigations are important to neurobiology, implicated in disease states (e.g., spinal spasticity, pathophysiology of pain). The results will be relevant to understanding how spinal neuron excitability is controlled by therapeutic agents that alter afferent transmission by acting at GABA receptors, and hence should foster eventual development of more efficacious pharmacological treatment of spinal cord injuries, pain states, and other pathologies. Whole-cell, cell-attached, and outside-out patch clamp recordings from cultured embryonic and adult human dorsal root ganglion (DRG) neurons will be used to define the electrophysiological/pharmacological properties of GABA receptors. The results of these investigations will be compared with data from molecular biological experiments designed to determine the subset of GABA receptor subunit-encoding mRNAs expressed by cultured embryonic and adult human DRG neurons. RT-PCR on cultures of DRG neurons will reveal the potential array of involved subunits. Based on conclusions drawn from these data, the candidate subunit combinations reproducing the properties of the novel GABA receptor will be identified using exogenous expression systems and antisense oligonucleotides. The reduced number of subunits potentially responsible for the observed properties reduced in this manner to a manageable number will allow performance of single-neuron RT-PCR to correlate expression of functional properties with expression of particular subunit mRNAs. These investigations will allow the applicants to specify the properties of a defined human GABA receptor with unique pharmacological properties within a native neuronal environment.

描述：该提案旨在研究电生理学/ GABA受体的药理特征和亚基组成 位于人类初级传入神经元上。 拟议的研究源于 最近的调查结果令人着迷，这些结果确定了这种培养 人类初级传入神经元表达具有独特和独特的GABA受体 迄今未描述的药理特性。 两者的联系 大型移植中心和国际公认的脊髓 创伤部门为申请人提供了人类神经元的访问。 三 具有专业知识的单独实验室在：（1）神经元细胞培养； （2） GABA受体的电生理学和药理学； （3）分子 神经递质受体的生物学将参与这些研究。 总体意图是阐明这些新型人类GABA的特性 通过使用复杂的电生理学的组合，受体 技术，药理学探针和分子生物学方法 定义通道/分子特性并确定GABA 人类感觉神经元中的受体亚基。 拟议的研究对神经生物学很重要，涉及 疾病状态（例如脊柱痉挛，疼痛的病理生理学）。 这 结果将与了解脊柱神经元的兴奋性有关 由通过表演改变传入传播的治疗剂控制 在GABA受体中，因此应最终促进更多 有效的药理学治疗脊髓损伤，疼痛状态， 和其他病理。 全细胞，细胞连接和外部外部贴片夹录制的记录 培养的胚胎和成年人类背根神经节（DRG）神经元将 用于定义电生理/药理特性 GABA受体。 这些调查的结果将与 分子生物学实验的数据旨在确定子集 由培养的胚胎和 成人人类DRG神经元。 RT-PCR关于DRG神经元培养的RT-PCR将揭示 潜在的涉及亚基。 根据这些结论 数据，候选亚基组合，重现了 新型GABA受体将使用外源表达系统鉴定 和反义寡核苷酸。 潜在的亚基数量减少 负责以这种方式降低的观察到的特性 可管理的数字将使单神经RT-PCR的性能 功能特性的表达与特定的表达 亚基mRNA。 这些调查将允许申请人指定 具有独特药理的定义人GABA受体的特性 本地神经元环境中的特性。