MULTIPLE SITE OPTICAL RECORDING OF MEMBRANE POTENTIAL

电位法探针是一种染料，当它结合到 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示剂。它的光学性质 随着电势的变化而线性变化，可用于 监测动作电位、突触电位或其他变化 膜电压一次可从多个位置，无需使用 多个电极。二十多年来，我们的实验室一直在开拓 使用电位法探头的技术，并开发了新的 用于细胞神经生理学的光学方法，包括高 用于跨膜多点光学记录的分辨率系统 电压(MSORTV)，能够监测膜电位的变化 一次从多达464个位点中分离出来。我们现在将应用这些技术 对于神经系统在三个层面上的整合的研究： 刺激偶联的神经末梢释放多肽的作用 S潜在侵袭一处高度分叉的神经末梢， 以及完整的对一个完整的 哺乳动物的神经网络。 首先，我们将使用光散射方法，结合荧光 钙指示剂和电压敏感染料来验证这一假设 触发的钙离子从血管内储存库中的释放 哺乳动物释放神经肽所必需的(并且 商店可能就是水泡本身。) 其次，我们将使用电位法来记录染料的作用 S对神经末梢干的潜在侵袭可能受 内在控制机制，包括时间模式 活性，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电压表来监测 哺乳动物单纯性神经中所有神经元的电活动 唯一可以通过光学手段进行分析的系统， 豚鼠回肠黏膜下神经丛，并改编一种新的分析 用于阐明任务的工具(引力变换) 完全是一个完整神经网络的整体行为。

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.