Synthetic design of an all-optical electrophysiology system

项目概要/摘要 光遗传学包括一系列广泛的工具和技术，涉及光的使用， 利用分子遗传工具，驱动和监测神经系统中特定类型的兴奋细胞的活动， 和心与传统的电生理技术相比，这些方法的侵入性要小得多， 同时监测和操纵多个部位的电活动的潜力。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， 对新疗法的发展有着深远的影响。尽管他们的承诺，目前的一代 与标准电生理学方法相比，光遗传学致动器的性能较差。而膜 在典型的电生理学实验中，电位可以在亚电极上改变数百毫伏， 毫秒的时间尺度上，目前产生的光激活离子通道能够驱动膜电位 在一毫秒内只增加几毫伏。该领域的许多前沿发展都集中在 修饰和再工程化天然存在的离子通道，但这些方法具有一些固有的缺点。 局限性。在此，我们建议开发一类新的合成探针，作为光激活致动器 用于以高时间和空间分辨率控制膜电位和离子浓度。采用 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 更有效的致动器具有驱动细胞膜电位所需的吞吐量。另外这些 化学离子载体可以与遗传编码的光激活探针组合， 灵活性.拟议的研究利用了合成化学家的专业知识（Schomaker教授，UW- 化学）和离子通道电生理学家（Chanda教授，威斯康星大学神经科学）。这两个具体目标将 重点是：a）光活性离子载体和离子载体的设计和合成，B） 这些设计者离子载体和离子载体的光学和传输性质。

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.