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）：在生命实验动物中操纵完整的神经回路中定义的神经元种群的需求对于研究脑疾病的研究迫切，特别是可能由异常电路引起的脑部疾病，例如许多经典精神疾病。长期目标是开发和完善体内神经元操作的更复杂的方法。该特定应用的总体目的是创建一组新的轻型技术，用于研究完整大脑中的神经电路动力学。这些技术基于将光遗传学与生物发光相结合，即将光感应分子（OPSINS）与通过荧光素酶进行生物学产生的光相结合。具有光生产和光感应的每个遗传编码的概念允许a）从标准的光遗传学应用中扩展Opsins的使用，还包括化学遗传激活； b）分别通过产前和突触后靶向光产生和光感应蛋白的突触信息的功能测试； c）通过钙敏感的荧光素酶将神经活动转化为光，将信号转换为光感应蛋白。拟议的研究的基本原理是，对电路的双峰询问，功能绘制长期和局部连通性以及使神经元的遗传靶向的非侵入性自我调节能够增加我们对神经元信息流的理解，从而有可能导致新的和改善神经精神疾病的方法。基于强大的初步数据，将通过追求三个特定目标来实现目标：1）确定最佳的荧光素酶 - 粘蛋白组合组合，以激活和抑制神经元活性； 2）在突触伙伴中表达细胞中产生的荧光素酶和光感应蛋白； 3）结合一个荧光素酶，该荧光素酶在神经元激活后用光感应质子泵产生灯光。在第一个目标下，已经有效的神经元激活剂将被优化并扩展到神经元沉默。在第二和第三目的下，将探索两个新颖的概念，特别是跨突触的光激活和神经元活性控制的光激活。这种方法是创新的 使用遗传编码的光源激活遗传编码的光转导分子。拟议的研究非常重要，因为它使得能够提出长期存在的问题，这些问题目前无法使用可用的技术解决，因此有望提高和扩展对神经元电路功能和功能障碍的理解。最终，这种增加的理解有可能告知新的治疗剂，这将有助于减少美国不断增长的心理健康问题。