MULTIPLE SITE OPTICAL RECORDING OF MEMBRANE POTENTIAL

某些物质，当结合到神经细胞膜上时，心脏和 骨骼肌、唾液腺泡和其他细胞的行为类似于分子 膜电位指示剂。它们的光学性质 分子，尤其是荧光和吸光度，呈线性变化。 具有潜力的时尚，因此可以用来监控行动 电位、突触电位或膜电压的其他变化 一次访问大量站点，而无需使用 电极。我们建议开发更敏感的探测器，以扩大 与它们的使用相关的技术，并使用这些分子 迄今用于光学记录膜电位的电压计 单个神经元的不可及区域，如神经内分泌轴突 终末、轴突和树突以及许多部位 同时，在神经元的小集合和电联会中， 以研究活动的空间和时间模式。 首先，我们打算使用基于计算机的多站点光学系统 跨膜电压(MSORTV)的记录，已经构建和 能够监测多达124个膜电位的变化 一次记录整个联会的电活动模式 (如腺体组织)，并研究真正简单的属性 神经系统--人工构建的突触小集合 在培养中保持的连接的无脊椎动物中枢神经元--由 从其所有组件以光学方式记录电活动 同时。 其次，我们建议使用这种装置，并使用一束氩离子激光 来源，以记录单个细微过程的膜电位变化 神经元在原位，在无脊椎动物神经管内，在组织中分离 文化。这些结构不能被微电极穿透，并且 经常太远，电的，他们的活动不能被反映出来 在躯体中。 最后，我们期望开发电位法的光学性质。 探头，以及我们的多点光学记录能力，以检测 脊椎动物神经末梢的电位变化及其相互关系 神经末梢动作电位形态的改变 神经肽的释放。

Certain substances, when bound to the membranes of neurons, cardiac and skeletal muscle, salivary acini, and other cells, behave as molecular indicators of membrane potential. The optical properties of these molecules, most notably fluorescence and absorbance, vary in a linear fashion with potential and may, therefore, be used to monitor action potentials, synaptic potentials, or other changes in membrane voltage from a large number of sites at once, without the necessity of using electrodes. We propose to develop more sensitive probes, to extend the technology associated with their use, and to employ these molecular voltmeters for optical recording of membrane potential from hitherto inaccessible regions of single neurons such as axon an neuroendocrine terminals and axonal and dendritic processes and from many sites simultaneously in small assemblages of neurons and in electrical syncitia, in order to study the spatial and temporal patterning of activity. First, we intend to use a computer based system for Multiple Site Optical Recording of Transmembrane Voltage (MSORTV), already constructed and capable of monitoring changes in membrane potential from as many as 124 loci at once, to record patterns of electrical activity throughout syncitia (such as glandular tissue), and to study the properties of truly simple nervous systems -- small artificially constructed ensembles of synaptically connected invertebrate central neurons maintained in culture -- by recording electrical activiy optically from all of their components simultaneously. Second, we propose to use this appratus, with an Argon ion laser light source, to record membrane potential changes from fine processes of single neurons in situ, within an invertebrate neuropil, and isolated in tissue culture. These structures are not penetrable by microelectrodes and are frequently too far away, electrically, for their activity to be reflected in the somata. Finally, we expect to exploit the optical properties of potentiometric probes, and our multiple site optical recording capability, to detect potential changes in vertebrate nerve terminals, and to correlate alterations in the shape of the nerve terminal action potential with the release of neuropeptides.