Comprehensive Structural and Functional Mapping of Mammalian Colonic Nervous System

摘要 结肠是许多疾病的所在地，这些疾病是导致发病率和死亡率的主要原因 巨大的经济负担。结肠内(肠)神经和外源性神经在调节中起着关键作用。 分泌运动、内分泌、免疫功能和痛感。人们越来越多地了解到 结肠肠道的神经化学和电生理特性、细胞生理学和功能作用 神经元及其与副交感、交感和感觉系统的相互作用 几十年。然而，这些数据主要是从小动物研究和相关知识中获得的 动物和人类都处于滞后状态，这阻碍了有效疗法的开发。最新进展 在包括3-D测绘在内的前沿方法中，创新的病毒跟踪工具和神经成像提供了 获取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， 这是以前从未实现过的。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图谱 在人类和猪中，作为一个相关的大型动物模型，基于其结构和生理上的相似性 人类。小鼠将被用于转基因、光遗传学和病毒追踪方法的研究。这 这一目标将通过开发出最先进技术的世界专家调查人员的共同努力来实现 神经解剖、分子、电生理和功能方法。获得的初步数据 该联盟的团队展示了使用Clarity、高分辨率共聚焦显微镜、病毒跟踪和 光遗传学，提供外源性神经纤维、肠道回路的详细图谱和探测的专业知识 人结肠肠神经元电生理学快速高分辨率神经成像。此外， 新的微电极阵列和光纤技术的设计使得量化电机模式成为可能 神经刺激达到了前所未有的分辨率水平。综合努力和多学科 这些方法将填补目前关于结肠内部和外部神经元回路和细胞-细胞的知识空白 交流，尤其是在人类组织和猪中，这是一种非常适合翻译的大型动物模型 对患者的适用性。这些发现将为理解这个器官中的神经回路和 对于潜在的电击干预治疗结肠疾病将是至关重要的。

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.