Determining the Role of Bacterial Products on Neuronal Localization and Function in a Symbiotic Organ

项目摘要 细菌，如在脊椎动物肠道微生物组中发现的那些，已被证明具有广泛的生物活性。 对许多动物的肠道（ENS）和中枢神经系统（CNS）均有影响。反过来， ENS已被证明对肠道中微生物组建立的进展有影响。然而，在这方面， 尽管细菌对神经元发育和生理学有着深远的影响， 脊椎动物肠道微生物组和ENS的复杂性使得这些现象背后的机制 很难研究。尽管主题和脊椎动物模型很重要，但这些系统涉及 微生物组在空间和组成上都很复杂，通常有数千种细菌， 可以在不同的宿主之间传播。因此，简单、精细可操作的模型系统 研究细菌共生体对神经元发育和功能的影响。解决 由于缺乏细菌影响宿主神经生物学机制的模型，我们建议使用 夏威夷短尾乌贼（Euprymna cumberopes）与其发光细菌共生体之间的共生关系 费氏弧菌作为研究细菌如何驱动宿主神经元发育的易处理系统。的 鱿鱼-弧菌系统是研究细菌产物对宿主生理学影响的成熟模型， 发展，其结果往往是普遍适用于其他动物系统。这项建议旨在 将鱿鱼-弧菌系统发展成为研究微生物对宿主神经生物学影响的新模型。 在我们的第一个目标中，我们将定义光器官的神经元景观，包括神经元的位置和身份 以及共生和细菌产物对两者的影响。为了实现这一目标，我们将开发新的方法 在我们的系统中进行RNA和蛋白质的双重标记，并采用神经元示踪技术进行成像， 轻风琴在我们的第二个目标中，我们提出研究神经肽信号转导在转导微生物中的作用， 从费氏弧菌到共生器官外部组织的信号。具体来说，我们将确定 在共生的前三天，FMRFamide信号在光器官中的位置和动力学， 研究FMRFamide信号传导对细菌介导的胚胎后发育的影响。数据 这一提议不仅将为一个强大的新模型系统奠定基础，而且将 为社区创造新的资源和技术，并就社区的作用提供新的见解。 外周神经系统在增强微生物介导的发育中的作用。

PROJECT SUMMARY Bacteria, such as those found in the vertebrate gut microbiome, have been shown to have widespread effects on both the enteric (ENS) and central nervous systems (CNS) in numerous animals. Reciprocally, the ENS has been shown to have an impact on the progression of microbiome establishment in the gut. However, despite the profound influence of bacteria on neuronal development and physiology, the diversity of the vertebrate gut microbiome and the complexity of the ENS make the mechanisms underlying these phenomena difficult to study. Despite the importance of the topic and vertebrate models, these systems involve microbiomes that are both spatially and compositionally complex, often with thousands of bacterial species that can very between individual hosts. Therefore there is a role for simple, exquisitely manipulable model systems in which to study the effects of bacterial symbionts on neuronal development and function. To address the dearth of such models for the mechanisms by which bacteria influence host neurobiology, we propose to use the symbiosis between the Hawaiian bobtail squid (Euprymna scolopes) and its luminescent bacterial symbiont Vibrio fischeri as a tractable system in which to study how bacteria drive host neuronal development. The squid-vibrio system is a well-established model for the effects of bacterial products on host physiology and development, with outcomes that are often generalizable to other animal systems. This proposal aims to develop the squid-vibrio system into a new model for the study of the effects of microbes on host neurobiology. In our first aim we will define the neuronal landscape of the light organ, including neuronal position and identity and the effect of symbiosis and bacterial products on both. To achieve this aim, we will develop new methods for dual labeling of RNA and proteins in our system and adapt neuronal tracing techniques for imaging in the light organ. In our second aim we propose to study the role of neuropeptide signaling on transducing microbial signals from V. fischeri to tissues on the outside of the symbiotic organ. Specifically, we will determine the location and dynamics of FMRFamide signaling in the light organ over the first three days of symbiosis and will examine the effects of FMRFamide signaling on bacterially-mediated post-embryonic development. The data arising from this proposal will not only lay the foundation for a powerful new model system, but will also generate new resources and techniques for the community and provide new insights as to the role of the peripheral nervous system in potentiating microbially-mediated development.