MULTIPLE SITE OPTICAL RECORDING OF MEMBRANE POTENTIAL

膜电位的多位点光学记录

• 批准号: 2445711
• 负责人: BRIAN Matthew SALZBERG
• 金额: $ 39.43万
• 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 1998-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2445711
• 关键词: Anura; Aplysia; alternatives to animals in research; axon; central nervous system; dendrites; fluorescent dye /probe; laboratory mouse; membrane potentials; neuropeptides; neuropil; online computer; potentiometry; synapses; tissue /cell culture

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.

某些两亲性染料，当结合到神经元的膜上时， 骨骼肌、腺体和其他细胞， 膜电位的指标。 这些的光学性质 分子随电位线性变化， 电位、突触电位或膜电压的其他变化， 大量的网站一次，而无需使用电极。 项目 将继续开发更灵敏的探针，增加我们对 他们降低电压的机制， 与他们的使用有关。 我们目前的目的是利用这些分子 光学记录功能性膜电位的电压表 硬骨鱼视网膜中的神经节细胞阵列，与其他情况不同 单个神经元不可及的区域，如神经末梢， 树枝状分支，并从许多网站同时在小 组织培养中的神经元集合。 首先，我们将使用新完成的高分辨率系统进行多个 跨膜电压的现场光学记录（MSORTV），能够 同时监测多达464个位点的膜电位变化， 识别和可视化视网膜神经节细胞内的功能单位， 当硬骨鱼的光感受器的双锥系统 用具有其电矢量的线偏振光估计 在两个正交方向之间快速交替。 这些实验 应该允许检测偏振对手视网膜神经节 细胞 其次，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 研究快速分泌细胞兴奋-分泌偶联的机制， 系统，并了解如何改变入侵的高度 单细胞神经元的分支终末分支可用于 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 从单个神经元的精细过程的变化，以确定 神经元结构的主动和被动电特性 其不能被电极穿透并且经常太远， 电，使它们的活动反映在胞体中。 最后， 我们将使用高分辨率多站点光学记录来继续我们的 研究活动的空间和时间模式，在真正简单的 神经系统，突触连接的小集合体， 通过监测所有神经元的电活动， 细胞同时