Mechanisms of Inhibitory Glycine Receptor Modulation

抑制性甘氨酸受体调节机制

• 批准号: 6623192
• 负责人: KWEE LIU LIN THIO
• 金额: $ 16.4万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-01 至 2007-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6623192
• 关键词: GABA receptor; Xenopus oocyte; barbiturates; chelating agents; colchicine; cytoskeleton; dithiol; electrodes; electrophysiology; gene targeting; genetically modified animals; glutathione; glycine receptors; hippocampus; human tissue; kidney cell; laboratory mouse; neurons; oxidation reduction reaction; pentobarbital; protein protein interaction; receptor expression; reducing agents; sulfhydryl reagents; tissue /cell preparation; voltage /patch clamp; zinc

K. Liu Lin Thio, MD, PhD is a pediatric epileptologist who is interested in developing a research career in ion channel modulation because of its importance to understanding and treating neurological diseases such as epilepsy. He has extensive experience with cellular neurophysiology but would like to probe the molecular mechanisms underlying ion channel modulation. This requires that he learn the basic techniques of molecular biology, which is one of the goals of this proposal. Several neurological disorders including epilepsy may result, in part, from cortical inhibitory glycine receptor (GlyR) dysfunction. Thus, GlyR modulation is important to understanding and treating neurological disease. Although several modulators of GlyR have been identified, their mechanisms of action are unknown because quantitative pharmacological and electrophysiological studies have not been performed. This study proposes to test three hypotheses regarding the mechanism by which three known GlyR modulators act: 1) GlyR and gamma-aminobutyric acidA (GABAA) receptors interact through the cytoskeleton; 2) Sulfhydryl reducing agents inhibit GlyR by chelating extracellular zinc; 3) Potentiation and inhibition of GlyR currents by barbiturates occur at distinct sites. These hypotheses will be tested by studying the electrophysiological properties of native GlyR in cultured embryonic mouse hippocampal neurons and GlyR expressed at Xenopus oocytes and human embryonic kidney (HEK) 293 cells.

K. Liu Lin Thio，MD，PhD是一位儿科癫痫病学家，他有兴趣在离子通道调节方面开展研究，因为它对理解和治疗癫痫等神经系统疾病非常重要。他在细胞神经生理学方面有着丰富的经验，但他想探索离子通道调节的分子机制。这需要他学习分子生物学的基本技术，这也是本提案的目标之一。包括癫痫在内的几种神经系统疾病可能部分由皮质抑制性甘氨酸受体（GlyR）功能障碍引起。因此，GlyR调节对于理解和治疗神经系统疾病是重要的。虽然已经鉴定了几种GlyR的调节剂，但由于尚未进行定量药理学和电生理学研究，因此其作用机制尚不清楚。本研究拟验证三种已知GlyR调节剂作用机制的三个假设：1）GlyR和γ-氨基丁酸A（GABAA）受体通过细胞骨架相互作用; 2）巯基还原剂通过螯合细胞外锌抑制GlyR; 3）巴比妥酸盐对GlyR电流的增强和抑制发生在不同的位点。这些假设将通过研究培养的胚胎小鼠海马神经元和表达在非洲爪蟾卵母细胞和人胚肾（HEK）293细胞的天然GlyR的电生理特性进行测试。