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

项目摘要 我们建议开发一种可长期植入的、全光学的光遗传神经接口，它可以 光学神经调节副交感神经或周围神经中单个神经轴突的侵袭性 系统。拟议的接口将有利于人类疾病和残疾的治疗 颈迷走神经支配的胸腹部器官和系统，如癫痫和 代谢紊乱。我们建议从这些神经的传入/传出轴突与 调节器官或由它们支配的大脑回路的目标。双向光神经接口 技术将利用通过病毒载体转染传入细胞实现的光遗传学能力 和/或带有基因靶向的、光学激活的报告蛋白和视蛋白的传出神经元。 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。这可以使用以下命令完成 基因编码的钙指示剂或其他改变其荧光的电压敏感蛋白 在神经元中产生动作电位时的特性。因为神经不会自然地表达光学 蛋白质，我们将与表达这些蛋白质的转基因小鼠合作，并使用这些小鼠模型来提炼 我们的系统在提供给其他研究人员使用之前。 我们的目标是开发一种紧凑的台式光学系统，可以与其他研究实验室共享，以提供 能够询问神经中特定的神经束和轴突的独特能力。在未来，这是 技术具有转化为人类临床应用的潜力。提案中的技术是 雄心勃勃，但我们已经组建了一支由细胞生物学家、神经学家、生物医学、电学、 和机械工程师。该团队拥有在多个项目上成功协作的良好记录 赠款和出版物。

Project Abstract We propose to develop a chronically implantable, all optical, optogenetic nerve interface that can non- invasively, optically neuromodulate individual axons of nerves in the parasympathetic or peripheral nervous system. The proposed interface would benefit treatment of human disease and disabilities related to the thoracic and abdominal organs and systems innervated by the cervical vagus nerve, such as epilepsy and metabolic disorders. We propose to optically interface from afferent/efferent axons in these nerves with the goal of modulating organs or brain circuits innervated by them. The bidirectional optical neural interface technology will utilize the capabilities of optogenetics enabled through viral vector transfection of afferent and/or efferent neurons with genetically targeted, optically activated reporter proteins and opsins. 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 genetically encoded calcium indicators or other voltage sensitive proteins that change their fluorescent properties upon action potential generation in a neuron. Because nerves do not naturally express optical proteins, we will work with transgenic mice that express these proteins and use these mice models to refine our system before making it available for other researchers to use. We aim to develop a compact, bench-top optical system that can be shared with other research labs to provide the unique ability of being able to interrogate specific fascicles and axons within the nerve. In the future, this technology has potential for translation to human clinical applications. The technology in the proposal is ambitious, but we have formed an outstanding team of cell biologists, neuroscientists, biomedical, electrical, and mechanical engineers. The team has an excellent track record of successful collaborations on multiple grants and publications.