Interrogating Neuronal Membrane Potential Dynamics with Optical Voltage Sensors

07项目摘要/摘要 这个项目的长期目标是在完整的背景下询问膜电压的动态 具有高时空分辨率的大脑。光学方法解剖神经元活动有望带来革命性的变化 我们在细胞和电路水平上对大脑的理解；然而，我们的理解仍然不完整 部分原因是缺乏能够以足够的速度和灵敏度直接报告神经元活动的工具。 我们建议利用分子设计和有机化学的力量来开发和应用新的光学工具 用于以前所未有的速度和灵敏度监测完整和非完整大脑的膜电位 与其他类别的电压指示器相关的破坏性电容性负载。我们计划利用光生技术 通过分子导线的电子转移(PET)作为光学电压传感的通用平台。我们将建造 一种用于光学电压传感的调色板，可延伸到电磁波的近红外区域 光谱；我们将创建新的电压传感器，具有非常高的双光子吸收截面 在厚厚的组织和完整的大脑中使用；我们将探索基因靶向和定位超 敏感的荧光电压传感器到感兴趣的神经元。在整个过程中，分子工具的开发将是 与神经元和组织中的应用密切相关，我们将应用这些工具来了解膜是如何 健康和神经疾病状态下的潜在动态变化。

07 Project Summary/Abstract The long-term goal of this project is to interrogate the dynamics of membrane voltage in the context of intact brains with high spatiotemporal resolution. Optical methods to dissect neuronal activity promise to revolutionize our understanding of the brain at the cellular and circuit level; however, our understanding remains incomplete due, in part, to a lack of tools that can report directly on neuronal activity with sufficient speed and sensitivity. We propose to use the power of molecular design and organic chemistry to develop and apply new optical tools for monitoring membrane potential with unprecedented speed and sensitivity in intact brains and without disruptive capacitive load associated with other classes of voltage indicators. We plan to exploit photoinduced electron transfer (PeT) through molecular wires as a versatile platform for optical voltage sensing. We will build a palette of colors for optical voltage sensing that extends into the near infrared regions of the electromagnetic spectrum; we will create new voltage sensors with exceptionally high two-photon absorption cross sections for use in thick tissue and intact brains; and we will explore methods for genetically targeting and localizing ultra- sensitive fluorescent voltage sensors to neurons of interest. Throughout, development of molecular tools will be closely wed to applications in neurons and tissues, and we will apply these tools to understand how membrane potential dynamics change in both healthy and neurological disease states.