SYNAPTIC FUNCTION--PHYSIOLOGY AND SUPERCOMPUTER STUDIES

突触功能——生理学和超级计算机研究

• 批准号: 2259947
• 负责人: JOEL R. STILES
• 金额: $ 7.32万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-09-30 至 1999-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2259947
• 关键词: acetylcholine; acetylcholinesterase; active sites; autoradiography; biophysics; computer simulation; disease /disorder model; electrophysiology; exercise; high voltage electron microscopy; intermolecular interaction; laboratory mouse; laboratory rat; lizards; molecular pathology; muscle cells; neuromuscular disorder; neuromuscular junction; neurotransmitter receptor; noise; synapses; temperature; voltage /patch clamp

These studies aim at reconstruction and detailed understanding of molecular events underlying microphysiological function at both normal and pathologically deranged synapses. Electrophysiological and ultrastructural investigations will be combined with supercomputer simulations employing novel Monte Carlo algorithms. At present, the focus is on the vertebrate neuromuscular junction and neuromuscular disease as a model system. In particular, experimental and simulation studies will address the biophysical factors critical to the generation of endplate currents at the normal junction, and the interplay of those factors in the pathogenesis and treatment of myasthenic syndromes. In the long run, methods and results obtained from these studies will be applied to other single and multi-synaptic systems in the peripheral and central nervous system. Present electrophysiological studies aim at determination of highly accurate endplate current data and single channel (acetylcholine receptor) data. Endplate current data will be obtained using highly optimized two electrode voltage clamp techniques. Single channel investigations will employ patch clamp and noise analysis techniques, which represent new training experience. Ultrastructural investigations will employ high voltage electron microscopic tomography for full scale three-dimensional reconstructions of junctional geometry. They will also employ quantitative electron microscopic autoradiography to determine synaptic densities of acetylcholinesterase in several preparations. The techniques to be employed in ultrastructural investigations also represent new training experience. All of the experimental information gained will serve as either input or test parameters for supercomputer simulations. Because of the use of Monte Carlo simulation algorithms, it will be possible to combine modeling of molecular acetylcholine diffusion, molecular interactions of acetylcholine with receptors and acetylcholinesterase, and full scale synaptic ultrastructure for the first time. This will yield important insights into synaptic function that were not previously obtainable.

这些研究旨在重建和详细了解 在正常和 病理性的突触。电生理和超微结构 调查将与使用超级计算机模拟合并 新颖的蒙特卡洛算法。目前，重点是脊椎动物 神经肌肉结和神经肌肉疾病作为模型系统。在 特别，实验和仿真研究将解决 生物物理因素对于在端板电流的产生至关重要 正常结，以及发病机理中这些因素的相互作用 和疗法综合征的治疗。从长远来看，方法和 从这些研究中获得的结果将应用于其他单个， 周围和中枢神经系统中的多突触系统。 目前的电生理研究旨在确定高度 准确的终板电流数据和单个通道（乙酰胆碱受体） 数据。终板电流数据将使用高度优化的两个 电极电压夹技术。单渠道调查将 采用贴片夹和噪声分析技术，代表新的 培训经验。超微结构调查将采用较高的 全尺度的电压电子微观层析成像 连接几何的重建。他们还将使用定量 电子显微镜放射自显影，以确定 乙酰胆碱酯酶在几项制剂中。这项技术是 从事超微结构调查也代表了新的培训 经验。获得的所有实验信息将作为 超级计算机模拟的输入或测试参数。由于 使用蒙特卡洛模拟算法，可以 结合了分子乙酰胆碱扩散，分子的模型 乙酰胆碱与受体和乙酰胆碱酯酶的相互作用，以及 首次全尺度突触超微结构。这将产生 对突触功能的重要见解 可获得。