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.

一种能激活/报告副交感神经轴突活动的光学探针 对于研究胰腺的研究人员来说，这一系统将是一个福音。我们正在提议开发这样一种探测器 利用我们在周围神经、生物成像和紧凑型光遗传学方面的背景和经验 多光子显微镜设计。目前迷走神经的神经调节方法通常都是 没有引起心率同时变化的事件，例如，与胰腺变化一起 功能。 我们建议开发一种新型的紧凑型光致发光型光学探头 在副交感神经或外周神经内对单个传入和/或传出轴突进行神经调节， 神经系统。我们寻求从这些神经读入或读出，目的是调节器官或大脑 它们所支配的回路。 我们的中心前提是，我们可以使用光学与神经中的轴突进行交流。对于光纤 工作方法我们需要将动作电位转换成光信号。这可以使用Report来完成 蛋白质或其他一些辅助动作电位产生的方式。因为神经不会 自然表达光学蛋白，我们将与表达这些蛋白的转基因小鼠合作，并使用这些 在提供给其他研究人员使用之前，让老鼠完善我们的系统。 我们将开发一种台式光学仪器，可以与其他研究团队共享，以允许我们， 以及它们，来询问老鼠，最终是人类神经中的特定神经束和轴突。我们的目标 这是胰腺的迷走神经及其神经支配。迷走神经是进入 副交感神经系统。与这种神经相互作用的能力使人有能力 神经调节内脏朝一个方向，大脑朝另一个方向。我们提议将一台光学元件 带有电润湿镜镜头的光纤，允许使用工作台远程询问迷走神经(即 便携式)激光系统。微型(直径1 mm)电润湿电可调光学元件的集成 基于光纤的成像系统将通过以下方式实现对神经元活动的双光子荧光成像 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、完善和展示我们的 从体外小鼠迷走神经激活/报告的能力，以及看看我们是否可以控制和/或感觉胰腺 在没有其他反应的情况下的反应，如心率变化，使用靶向神经调节 在体内转基因小鼠实验中迷走神经中特定轴突的研究。