MULTIPLE SITE OPTICAL RECORDING OF MEMBRANE POTENTIAL

膜电位的多位点光学记录

• 批准号: 2891597
• 负责人: BRIAN Matthew SALZBERG
• 金额: $ 35.69万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-12-01 至 2001-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2891597
• 关键词: Xenopus; calcium flux; endoplasmic reticulum; exocytosis; gamma aminobutyrate; guinea pigs; ileum; laboratory mouse; laboratory rat; membrane potentials; neural information processing; neural transmission; neurohypophysis; potentiometry; tissue /cell culture; visceral afferent nerve

Potentiometric probes are dyes which, when bound to the membranes of neurons, cardiac and skeletal muscle, glands, and other cells, behave as molecular indicators of membrane potential. The optical properties of these molecules vary linearly with potential and may be used to monitor action potentials, synaptic potentials or other changes in membrane voltage from a large number of sites at once, without the use of electrodes. For more than twenty years our laboratory has pioneered the technology for using potentiometric probes, and developed new optical methods for use in cellular neurophysiology, including a high resolution system for Multiple Site Optical Recording of Transmembrane Voltage (MSORTV), capable of monitoring changes in membrane potential from as many as 464 loci at once. We will now apply these techniques to the study of the nervous system at three levels of integration: the stimulus-coupled release of peptides from nerve terminals, the action potential s invasion of a highly ramified nerve terminal arborization, and the complete analysis of the ensemble behavior of an intact mammalian neural network. First, we will use light scattering methods, together with fluorescent calcium indicators and voltage sensitive dyes to examine the hypothesis that the triggered release of calcium from intraterminal stores is required for the release of neuropeptides in mammals (and that the stores may be the vesicles themselves.) Second, we will use potentiometric dyes to record how the action potential s invasion of a nerve terminal arbor may be modulated by intrinsic control mechanisms, including the temporal patterning of activity, and the local release of an inhibitory neurotransmitter (gamma-aminobutyric acid). Finally, we will use these molecular voltmeters to monitor the electrical activity of all of the neurons in a mammalian simple nervous system that is uniquely amenable to analysis by optical means, the submucous plexus of the Guinea-pig ileum, and adapt a novel analytical tool (the gravitational transformation) to the task of elucidating completely the ensemble behavior of an intact neural network.

电位探针是染料，当结合到细胞膜上时， 神经元、心肌和骨骼肌、腺体和其他细胞， 作为膜电位的分子指标。 光学性质 这些分子的浓度随电位线性变化， 监测动作电位，突触电位或其他变化， 膜电压从大量的网站一次，而无需使用 的电极。 二十多年来，我们的实验室一直是 使用电位探针的技术，并开发了新的 用于细胞神经生理学的光学方法，包括高 用于跨膜的多位点光学记录的分辨率系统 电压（MSORTV），能够监测膜电位的变化 464个基因座的基因 我们现在将应用这些技术 神经系统的研究在三个层次的整合： 刺激偶联释放肽从神经末梢，行动 高度分叉的神经末梢分支的潜在侵袭， 以及完整的整体行为分析 哺乳动物神经网络 首先，我们将使用光散射方法，连同荧光 钙指示剂和电压敏感染料来检验假设 终末内钙储存的触发释放 在哺乳动物中释放神经肽所需的（并且， 储存物可以是囊泡本身。） 其次，我们将使用电位染料来记录如何行动 电位对神经末梢的侵入可以通过 内在的控制机制，包括时间模式 活动，以及抑制性神经递质的局部释放 （γ-氨基丁酸）。 最后，我们将使用这些分子伏特计来监测 哺乳动物简单神经系统中所有神经元的电活动 系统是唯一适合通过光学手段分析， 豚鼠回肠粘膜下神经丛，并采用一种新的分析 工具（引力变换）来阐明 完整的神经网络的整体行为。