Synthetic design of an all-optical electrophysiology system

项目摘要/摘要 光遗传学包括一系列广泛的工具和技术，这些工具和技术涉及联合使用光 利用分子遗传学工具，驱动和监测神经系统中特定类型的可兴奋细胞的活动 还有心。与传统的电生理技术相比，这些方法侵入性小得多，而且 同时监控和操控多个地点的电子活动的潜力。的承诺 光遗传学不仅限于扩展我们对复杂器官系统的基本理解，还将 对新疗法的发展产生了深远的影响。尽管他们承诺过，但当前这一代人 与标准的电生理方法相比，光遗传致动器的性能较差。而膜 在一个典型的电生理实验中，电压可以在几百毫伏以下改变 毫秒级的时间尺度，当前一代的光激活离子通道能够驱动膜电位 每毫秒只有几毫伏。该领域的许多尖端发展都集中在 修改和重新设计自然产生的离子通道，但这些方法有一些固有的 限制。在这里，我们建议开发一类新的合成探针，作为光激活的执行器 用于控制高时间和空间分辨率的膜电位和离子浓度。用人 对这些探针的化学合成方法将使我们能够更灵活地设计和设计 更高效的致动器，具有驱动细胞膜电位所需的吞吐量。此外，这些 化学离子载体可以与基因编码的光活化探针相结合，以提供更大的 灵活性。这项拟议的研究利用了一位合成化学家(威斯康星大学舒梅克教授)的专业知识。 一位离子通道电生理学家(威斯康星大学神经科学学院昌达教授)。这两个具体目标将 重点是：a)设计和合成光活性离子载体和离子载体，b)表征 这些设计的离子载体和离子载体的光学和传输特性。

Project Summary/Abstract Optogenetics encompasses a broad array of tools and techniques that involve the use of light, in conjunction with molecular genetic tools, to drive and monitor activity of specific types of excitable cells in the nervous system and heart. Compared to traditional electrophysiological techniques, these methods are far less invasive and have the potential to monitor and manipulate electrical activity at multiple sites at the same time. The promise of optogenetics is not solely limited to expanding our basic understanding of complex organ systems but will also have a profound impact on the development of new therapeutics. Despite their promise, the current generation of optogenetic actuators are inferior compared to standard electrophysiological methods. While the membrane potential in a typical electrophysiological experiment can be changed by hundreds of millivolts on a sub- millisecond timescale, the current generation of light-activated ion channels are able to drive membrane potential by only a few millivolts in a millisecond. Much of the cutting-edge development in the field has focused on modifying and reengineering naturally-occurring ion channels, but these approaches have some inherent limitations. Herein, we propose to develop a new class of synthetic probes that serve as light-activated actuators for controlling membrane potential and ion concentrations with high temporal and spatial resolution. Employing a chemical synthesis approach towards these probes will allow us much greater flexibility to engineer and design more efficient actuators having the necessary throughput to drive cellular membrane potential. In addition, these chemical ion carriers can be combined with genetically encoded light-activated probes to provide even greater flexibility. The proposed research capitalizes on the expertise of a synthetic chemist (Prof. Schomaker, UW- Chemistry) and an ion channel electrophysiologist (Prof. Chanda, UW-Neuroscience). The two specific aims will focus on: a) the design and synthesis of photoactive ionophores and ion carriers, b) Characterization of the optical and transport properties of these designer ionophores and ion carriers.