Genetically Encoded Light-Production and Light-Sensing for Neuronal Manipulation

用于神经元操纵的基因编码光产生和光传感

• 批准号: 8971048
• 负责人: UTE H HOCHGESCHWENDER
• 金额: $ 17.33万
• 依托单位: CENTRAL MICHIGAN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-12-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8971048
• 关键词: Genetically; Encoded; Light; Production; Sensing

DESCRIPTION (provided by applicant): The need to manipulate defined neuronal populations in intact neural circuits in the living experimental animal is urgent for the study of brain disorders, specifically those which are likely caused by abnormal circuitry, such as many of the classical psychiatric disorders. The long-term goal is to develop and refine ever more sophisticated methods for in vivo neuronal manipulation. The overall objective of this particular application is to create a set of new light-driven technologies for studying neural circuit dynamics in the intact brain. These technologies are based on combining optogenetics with bioluminescence, i.e. combining light-sensing molecules (opsins) with biologically produced light through luciferases. The concept of having light-production and light-sensing each genetically encoded permits a) to expand the use of opsins from standard optogenetic applications to also include chemical genetic activation; b) functional testing of synaptic information flow through pre- and postsynaptic targeting of the light-producing and the light-sensing proteins, respectively; and c) translating neural activity via calcium-sensitive luciferases into light whichin turn signals to a light-sensing protein. The rationale for the proposed research is that bimodal interrogation of circuits, functionally mapping long-range and local connectivity, and enabling genetically targeted non-invasive self-regulation of neurons will increase our understanding of neuronal information flow, potentially resulting in new and improved approaches to treatment of neuropsychiatric disorders. Based on strong preliminary data, the objectives will be achieved by pursuing three specific aims: 1) Identify the optimal luciferase-opsin combinations for activating and inhibiting neuronal activity; 2) Express the light-producing luciferase and the light-sensing opsin in cells across synaptic partners; and 3) Combine a luciferase which produces light upon neuronal activation with a light-sensing proton pump. Under the first aim, already working versions of neuronal activators will be optimized and extended to neuronal silencing. Under the second and third aims two novel concepts will be explored, specifically transsynaptic light activation and neuronal activity-controlled light activation. The approach is innovative in that it uses a genetically encoded light source to activate a genetically encoded light transducing molecule. The proposed research is significant, because it enables to ask longstanding questions which currently cannot be addressed with available technology and thus is expected to advance and expand understanding of neuronal circuit function and dysfunction. Ultimately, such increased understanding has the potential to inform new therapeutics that will help reduce the growing mental health problems in the United States.

描述（由申请人提供）：为了研究脑部疾病，特别是那些可能由异常电路引起的大脑疾病，例如许多经典精神疾病，迫切需要在活体实验动物的完整神经回路中操纵已定义的神经元群。长期目标是开发和完善更复杂的方法，用于体内神经元操作。这项特殊应用的总体目标是创造一套新的光驱动技术，用于研究完整大脑中的神经回路动力学。这些技术基于光遗传学与生物发光的结合，即通过荧光素酶将光感分子（视蛋白）与生物产生的光结合起来。每个基因编码的光产生和光感应的概念允许a)将视蛋白的使用从标准的光遗传学应用扩展到还包括化学遗传激活；B)分别通过对产光蛋白和光感蛋白的突触前靶向和光感蛋白的突触后靶向来检测突触信息流的功能；c)通过钙敏感的荧光素酶将神经活动转化为光，进而向光感应蛋白发出信号。提出这项研究的基本原理是，电路的双峰询问，远程和局部连接的功能映射，以及实现神经元的基因靶向非侵入性自我调节，将增加我们对神经元信息流的理解，可能导致新的和改进的神经精神疾病治疗方法。基于强大的初步数据，目标将通过追求三个具体目标来实现：1)确定激活和抑制神经元活动的最佳荧光素酶-视蛋白组合；2)在突触伴侣细胞中表达产光荧光素酶和光感视蛋白；3)结合在神经元激活时产生光的荧光素酶和光感应质子泵。在第一个目标下，已经工作的神经元激活器将被优化并扩展到神经元沉默。在第二个和第三个目标下，将探索两个新概念，特别是跨突触光激活和神经元活动控制的光激活。这种方法是创新的