Development of a Bidirectional Optogenetic Minimally Invasive Peripheral Nerve Interface with Single Axon Read-in & Read-out Specificity

单轴突读入双向光遗传学微创周围神经接口的开发

• 批准号: 9535582
• 负责人: RICHARD Fergus ffrench WEIR
• 金额: $ 64.52万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-24 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9535582
• 关键词: Action Potentials; Afferent Neurons; Alpha Cell; Axon; Beta Cell; Biological; Brain; Breeding; Caliber; Cardiac; Cervical; Collaborations; Coupled; Detection; Development; Diabetes Mellitus; Diabetic Autonomic Neuropathy; Efferent Neurons; Event; Fascicle; Fiber Optics; Fluorescent Dyes; Generations; Glucagon; Goals; Head; Heart; Heart Rate; Hormones; Human; Image; Implant; In Vitro; Individual; Insulin; Islets of Langerhans; Label; Langerhans cell; Lasers; Measures; Methods; Microscope; Microscopy; Mus; Muscle fasciculation; Nerve; Neurons; Neurosciences; Optical Instrument; Optical reporter; Optics; Organ; Pancreas; Pancreatic Diseases; Parasympathetic Nervous System; Pathway interactions; Peripheral Nerves; Peripheral Nervous System; Physiological; Preparation; Proteins; Ranvier' s Nodes; Reporter; Reporting; Research; Research Personnel; Scanning; Sensory; Signal Transduction; Site; Source; Specificity; Stimulus; System; Technology; Testing; Tissues; Transgenic Mice; Vagus nerve structure; Viral Vector; Viscera; Work; aerobic respiration control protein; base; bioimaging; cholinergic; design; experience; experimental study; fluorescence imaging; imaging system; in vivo; instrument; insulin secretion; islet; lens; member; minimally invasive; nerve supply; neuroregulation; novel; optical fiber; optical imaging; optogenetics; pancreatic juice; portability; protein expression; response; rhodamine dextran; tool; two-photon

An Optical Probe capable of Activating/Reporting on axon activity in nerves of parasympathetic nervous system would be a boon to researchers working with the pancreas. We are proposing to develop such a Probe using our background and experience in optogenetics in the peripheral nerve, bio-imaging and compact multiphoton microscope design. Current neuro-modulation approaches for the vagus nerve are generally all or nothing events that cause simultaneous changes in heart rate, for example, along with changes in pancreatic function. We propose to develop a novel compact Optogenetic based Optical Probe capable of optically neuromodulating individual afferent and/or efferent axons within nerves of the parasympathetic, or peripheral, nervous system. We seek to read-in or read-out from these nerves with the goal of modulating organs or brain circuits innervated by them. Our central premise is that we can use optics to communicate with axons in a nerve. For optical approaches to work we need to convert action potentials into an optical signal. This can be done using reporter proteins or by some other means that is ancillary to action potential generation. Because nerves do not naturally express optical proteins, we will work with transgenic mice that express these proteins and use these mice to refine our system before making it available for other researchers to use. We will develop a bench-top Optical instrument that can be shared with other research teams to allow us, and them, to interrogate specific fascicles and axons within mouse, and ultimately human, nerves. Our goal here is the vagus nerve and its innervation of the pancreas. The vagus nerve is one of the main conduits into the parasympathetic nervous system. The ability to interface with this nerve gives one the ability to neuromodulate the viscera in one direction and the brain in the other. We are proposing to couple an optical fiber with an electrowetting lens head to allow remote interrogation the vagus nerve with a bench top (i.e. portable) laser system. Integration of miniature (1mm diameter) scale electrowetting electrically tunable optics with an optical fiber-based imaging system will enable two-photon fluorescence imaging of neuron activity by readout of a fluorescent indicator. We will work with our collaborators in the field of pancreatic research to test, refine and demonstrate our ability to activate/report from in-vitro mouse vagus nerves and to see if we can control and/or sense pancreatic responses in the absence of other responses, such as a change in heart rate, using targeted neuro-modulation of specific axons in the vagus in in-vivo transgenic mice experiments.

一种能够激活/报告副交感神经神经中轴突活性的光学探针 系统将为与胰腺合作的研究人员带来福音。我们建议开发这样的探测 利用我们在外围神经，生物成像和紧凑的光遗传学方面的背景和经验 多光子显微镜设计。迷走神经的当前神经调节方法通常是全部或 例如，没有任何会导致心率同时变化的事件以及胰腺变化 功能。 我们建议开发一种新型紧凑的基于光遗传学的光学探针，能够光学上 神经调节在副交感神经或周围的神经内的个体传入和/或传出轴突 神经系统。我们寻求从这些神经中读取或读取，以调节器官或大脑的目标 他们支配的电路。 我们的中心前提是，我们可以使用光学元件与神经的轴突进行通信。用于光学 工作方法我们需要将动作电位转换为光学信号。这可以使用记者完成 蛋白质或某些其他手段是行动潜在产生的。因为神经没有 自然表达光蛋白，我们将与表达这些蛋白质的转基因小鼠一起工作并使用这些蛋白 在使其他研究人员使用之前，小鼠可以完善我们的系统。 我们将开发一种可以与其他研究团队共享的基准光学仪器，以允许我们 和它们，询问小鼠内部的特定束和轴突，最终是人类的神经。我们的目标 这是迷走神经及其胰腺的神经支配。迷走神经是主要导管之一 副交感神经系统。与这种神经接触的能力使一个能力 神经调节内脏沿一个方向和另一个方向进行神经调节。我们建议夫妇光学 带有电镜头头的纤维可允许远程询问带有长凳顶部的迷走神经（即 便携式）激光系统。微型（直径1毫米）的整合电气表电气可调光学元件 使用基于光纤的成像系统将通过 荧光指示器的读数。 我们将与胰腺研究领域的合作者合作测试，完善和证明我们的 能够激活/报告中的Vitro小鼠迷走神经的能力，并查看我们是否可以控制和/或感官胰腺 在没有其他反应的情况下，响应（例如，心率变化）使用靶向神经调节 体内转基因小鼠实验中迷走神经中的特定轴突的。