MULTIPLE SITE OPTICAL RECORDING OF MEMBRANE POTENTIAL

某些两亲性染料，当结合到神经元膜上时，心脏 和骨骼肌、腺体和其他细胞，表现为分子 膜电位指示剂。它们的光学性质 分子随电势呈线性变化，可用于监测行动。 电位、突触电位或膜电压的其他变化 大量的站点一次完成，无需使用电极。该项目 将继续开发更敏感的探测器，增加我们对 他们转换电压和扩展技术的机制 与它们的使用有关。我们目前的意图是利用这些分子 光学记录功能膜电位的电压表 硬骨鱼视网膜中的神经节细胞阵列 单个神经元的不可触及的区域，如神经末梢和 树枝分枝，并同时从多个地点在小 组织培养中的神经元集合。 首先，我们将使用我们新完成的高分辨率系统对多个 跨膜电压现场光学记录)，能够 同时监测多达464个基因座的膜电位变化， 识别和可视化视网膜神经节细胞内的功能单位 当硬骨鱼的双锥体感光系统 用具有其电矢量的线偏振光估计的 在两个垂直方向之间快速交替的。这些实验 应允许检测到与极化相反的视网膜神经节 细胞。第二，我们将利用实验室的监测能力 脊椎动物完整神经末梢的电活动顺序 快速分泌中兴奋-分泌耦合机制的研究 系统，并了解如何在高度入侵的变化 单细胞神经元的分枝终末分支可用于 调节哺乳动物神经垂体中神经肽的释放。 第三，新的MSORTV系统将用于记录膜电位 从单个神经元细小突起的变化来确定 神经元结构的有源和无源的电学特性 它们不能被电极穿透并且经常太远， 在电学上，它们的活动被反映在体细胞中。最后， 我们将使用高分辨率多站点光学记录来继续我们的 《真简单》中活动的时空模式研究 神经系统，突触连接的中枢海兔的小集合 在培养中维持的神经元，通过监测所有细胞的电活动 它们的细胞同时存在。

Certain amphipathic dyes, 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 site at once, without the use of electrodes. The project will continue to develop more sensitive probes, add to our understanding of the mechanisms by which they transduce voltage, and extend the technology associated with their use. Our present intention is to use these molecular voltmeters for optical recording of membrane potential from functional arrays of ganglion cells in retinae of teleost fishes, from otherwise inaccessible regions of single neurons such as nerve terminals and dendritic arborizations, and from many sites simultaneously in small ensembles of neurons in tissue culture. First, we will use our newly completed 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 one, to identify and visualize functional units within the retinal ganglion cell mosaic of bony fishes when their double cone system of photoreceptors is estimated with linearly polarized light having its electric vector alternating rapidly between two orthogonal directions. These experiments should permit the detection of polarization-opponent retinal ganglion cells. Second, we will exploit the laboratory's ability to monitor electrical activity in the intact nerve terminals of vertebrates in order to study the mechanism of excitation-secretion coupling in a fast secretory system, and also to understand how change in the invasion of the highly ramified terminal arborization of monocellular neurons can be used to modulate the release of neuropeptides in the neurohypophyses of mammals. Third, the new MSORTV system will be used to record membrane potential changes from fine processes of single neurons in order to determine the electrical properties , both active and passive, of neuronal structures which are not penetrable by electrodes and are frequently too far away, electrically, for their activity to be reflected in the somata. Finally, we will use high resolution multiple site optical recording to continue our study of the spatial and temporal patterning of activity in truly simple nervous systems, small ensembles of synaptically connected Aplysia central neurons maintained in culture, by monitoring electrical activity in all of their cells simultaneously.