Functional mapping of enteric-associated neurons

肠神经系统包括胃肠道内复杂且广泛的网络， 其特征在于，在神经元的内部和外部都有包含神经元体的内在臂和外在臂。 肠，分别。与它们在肠道生理学中的作用一致，EAN功能受损可导致 与分泌和运动功能缺陷相关的病理学和慢性炎性病症。EANs 使肠组织与大量免疫细胞共存，免疫细胞和神经细胞都 其配备有监测腔表面处的扰动的感测机构。双向交互作用 免疫细胞和神经细胞之间的关系已经在稳定状态下被记录， 已经提出相互作用是几种疾病过程的一部分，既包括局部的（例如肠易激综合征）， 综合征）和系统性（例如多发性硬化症）。尽管它与人类生理学相关，但它所起的作用 通过EAN在组织维护和病理学中的作用，或EAN如何将管腔损伤传递到局部或远处 组织仍不清楚。获得CNS内神经元群体遗传通路的新方法 强调了这些技术的变革潜力。令人惊讶的是， 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EAN。通过结合新型成像和转录组学工具，我们的实验室开发了广泛的 在理解粘膜和肠道免疫反应方面的经验，我们最近的工作强调了 EAN在协调免疫反应中的作用。例如，我们发现了一个意想不到的角色 对于EAN通过外源性交感神经源性神经元调节结构偶联的巨噬细胞群体， 通过肠巨噬细胞上的肾上腺素能受体β 2（β2AR）的去甲肾上腺素信号传导。克服 在上述EAN研究中的障碍，该建议结合了细胞特异性 主动翻译核糖体分析、组织清除、神经元的光遗传和化学遗传调节 功能以及病毒追踪，以生成EAN的第一个功能图谱，定义微生物 肠道中的感应回路这里提出的实验不仅将映射该感测电路，而且还将映射该感测电路。 建立操纵EAN活性的工具，以构建EAN响应管腔挑战的功能图。 由于我们还建议使用这些新技术使用人类肠道样本，三个- 小鼠和人类样本之间的维度比较不仅会产生对进化的见解， 保守的机制和途径相关的EAN架构和行为，但也增加了一个强大的 翻译的组成部分，了解人类肠道生理和病理。拟建项目 因此，将提供一个急需的平台，以了解和探索新的治疗策略， 治疗与炎症诱导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.