Comprehensive Structural and Functional Mapping of Mammalian Colonic Nervous System

摘要 结肠是多种疾病的发生部位，这些疾病是发病率和死亡率的主要原因， 沉重的财政负担。结肠的内源性（肠）和外源性神经支配在调节 分泌运动、内分泌、免疫功能和痛觉。人们越来越了解 神经化学和电生理学特性，细胞生理学，以及结肠肠道的功能作用， 神经元及其与副交感神经，交感神经和感觉系统在过去的相互作用 几十年然而，这些数据主要来自小动物研究和大规模的相关知识。 动物和人类的研究滞后，这阻碍了有效疗法的发展。最新进展 包括3D绘图、创新的病毒追踪工具和神经成像在内的尖端方法， 获得大型动物和人体组织中神经回路和相关功能的详细信息的手段， 这是前所未有的。该提案的总体目标是提供一个全面和 结肠不同区域的内在和外在神经支配的详细结构和功能映射 在人类和猪中，作为一个相关的大型动物模型，基于其结构和生理相似性， 人类小鼠将用于涉及转基因、光遗传学和病毒追踪方法的研究。这 这一目标将通过世界专家研究人员的共同努力来实现，他们开发了最先进的 神经解剖学、分子学、电生理学和功能方法。获得的初步数据 该联盟的团队展示了使用高分辨率共聚焦显微镜，病毒追踪和 光遗传学提供详细的外部神经纤维，肠电路和专业知识，以探测 人结肠肠神经元电生理学研究此外该 新的微电极阵列和光纤技术的设计允许量化响应的运动模式 以前所未有的分辨率刺激神经。联合努力和多学科 这些方法将填补目前关于结肠内在和外在神经元回路以及细胞-细胞回路的知识空白。 通信，特别是在人类组织和猪，一个大型动物模型，非常适合翻译 适用于患者。这些发现将为理解该器官的神经回路奠定基础， 将是治疗结肠疾病的潜在电介入的关键。

ABSTRACT The colon is the site of a multitude of disorders which are leading causes of morbidity and mortality and of significant financial burden. The colonic intrinsic (enteric) and extrinsic innervation play crucial role in regulating the secreto-motor, endocrine, immune functions and pain sensation. There has been increasing understanding of the neurochemical and electrophysiological properties, cell physiology, and functional roles of colonic enteric neurons and their interaction with the parasympathetic, sympathetic and sensory systems during the past decades. However, these data are derived largely from small animal studies and relevant knowledge in large animals and humans is lagging, which has hampered the development of effective therapies. Recent advances in cutting edge approaches including 3-D mapping, innovative viral tracing tools, and neuroimaging provide a means to obtain detailed information on neural circuits and related functions in large animals and human tissues, which has never been achieved before. The overall objective of the proposal is to provide a comprehensive and detailed structural and functional mapping of the intrinsic and extrinsic innervation of various regions of the colon in humans and the pig, as a relevant large animal model based on its structural and physiological similarities to humans. Mice will be utilized for studies involving transgenic, optogenetics and viral tracing approaches. This objective will be achieved by a concerted effort of world expert investigators who developed state-of-the art neuroanatomical, molecular, electrophysiological and functional approaches. Preliminary data obtained by the consortium team showed the feasibility to use CLARITY, high resolution confocal microscopy, viral tracing and optogenetics to provide detailed mapping of extrinsic nerve fibers, enteric circuitries and the expertise to probe human colonic enteric neurons electrophysiologically by fast and high resolution neuroimaging. In addition, the design of new microelectrode array and fiber optic technology has allowed quantifying motor patterns in response to nerve stimulation at a resolution level not attained before. The combined effort and multidisciplinary approaches will fill the gaps in current knowledge on colonic intrinsic and extrinsic neuronal circuits and cell-cell communication, especially in human tissues and pig, a large animal model ideally suited for translational applicability to patients. These findings will set the foundation for understanding neurocircuitry in this organ and will be critical for potential electroceutical interventions to treat colonic disorders.