Functional mapping of enteric-associated neurons

肠道神经系统由胃肠道内复杂而广泛的网络组成， 其特征是内在和外在的臂都含有 肠，分别为。与它们在肠道生理中的作用一致，EAN功能受损可导致 与分泌和运动功能缺陷以及慢性炎症相关的病理。伊恩斯 肠道组织与大量免疫细胞共存，免疫细胞和神经元细胞都是 配备了监测管腔表面扰动的传感装置。双向互动 免疫细胞和神经细胞之间的关系已被证明处于稳定状态和功能障碍 相互作用被认为是几种疾病过程的一部分，既有局部的(例如肠易激) 综合症)和全身性(如多发性硬化症)。尽管它与人类生理学有关，但它扮演的角色 由eans在组织维护和病理学方面或eans如何向本地或远程传递管腔侮辱 组织仍不清楚。获得中枢神经内神经元群体遗传通路的新方法 突出了这些技术的变革潜力。令人惊讶的是，在这方面取得的进展很少 对中枢神经系统的研究已经转化为对周围神经系统的重要理解 包括Eans在内。通过结合新的成像和转录工具，我们的实验室开发了广泛的 了解粘膜和肠道免疫反应的经验，我们最近的工作突出了 Eans在协调免疫反应中扮演的角色。例如，我们发现了一个意想不到的角色 EANS通过外源性交感神经细胞来源调节结构耦合的巨噬细胞群 肠巨噬细胞上去甲肾上腺素通过肾上腺素能受体β2(β2AR)的信号转导。要克服 在上面提到的eANS研究中的障碍，这一建议结合了细胞特异性的最新进展 主动翻译核糖体轮廓、组织清除、神经元的光和化学遗传调制 功能以及病毒跟踪，以生成EAN的第一个功能图，定义微生物 肠道中的感应回路。这里提出的实验不仅将绘制这种传感电路，而且还将 建立操纵EAN活动的工具，以构建EAN的功能图，以应对重大挑战。 由于我们还计划使用这些使用人类肠道样本的新技术，这三项技术- 老鼠和人类样本之间的尺寸比较将不仅产生对进化的洞察 与EAN架构和行为相关的保守机制和路径，但也增加了强大的 了解人类肠道生理和病理的翻译部分。拟议中的项目 从而将提供一个急需的平台，以了解和探索新的治疗策略 治疗与炎症引起的神经元功能障碍相关的疾病。

The enteric nervous system comprises a complex and widespread network within the gastrointestinal tract and is characterized by both intrinsic and extrinsic arms containing neuron bodies within and outside of the intestine, respectively. Consistent with their role in gut physiology, impaired EAN function can lead to pathologies associated with defective secretory and motor function and chronic inflammatory conditions. EANs cohabitate the intestinal tissue with large populations of immune cells and both, immune and neuronal cells are equipped with sensing mechanisms that monitor perturbations at the luminal surface. Bidirectional interactions between immune and neuronal cells have been documented at steady state and dysfunction in these interactions have been proposed to be part of several disease processes, both local (e.g. irritable bowel syndrome) and systemic (e.g. multiple sclerosis). Despite its relevance for human physiology, the role played by EANs in tissue maintenance and pathology or how EANs communicate luminal insults to the local or distant tissues remains unclear. Novel approaches to gain genetic access to neuronal populations within the CNS have highlighted the transformative potential of these techniques. Surprisingly, little of the progress made in the study of the CNS has been translated into a significant understanding of the peripheral nervous system including EANs. By combining novel imaging and transcriptomic tools, our lab has developed extensive experience in understanding mucosal and intestinal immune responses, and our recent work has highlighted the role that EANs play in orchestrating immune responses. For instance, we uncovered an unexpected role for EANs in modulating a structurally coupled macrophage population via extrinsic sympathetic neuron-derived norepinephrine signaling through adrenergic receptor beta 2 (β2AR) on gut macrophages. To overcome obstacles in the study of EANs mentioned above, this proposal incorporates recent advances in cell-specific actively translating ribosome profiling, tissue clearing, opto- and chemo-genetic modulation of neuronal function as well as viral tracing in order to generate the first functional mapping of EANs, defining microbial sensing circuits in the intestine. Experiments proposed here will not only map this sensing circuit, but also establish tools to manipulate EAN activity to build a functional map of EANs in response to luminal challenges. Since we also propose to employ these novel techniques using human intestinal samples, the three- dimensional comparisons between mouse and human samples will yield insights not only into evolutionarily conserved mechanisms and pathways relevant to EAN architecture and behavior, but also add a strong translational component for understanding human intestinal physiology and pathology. The proposed project will thereby provide a much-needed platform to understand and explore novel therapeutic strategies for the treatment of disorders associated with inflammation-induced neuronal dysfunction.