Functional Integration of Newborn Olfactory Sensory Neurons in the Healthy and Regenerating Olfactory System

新生儿嗅觉感觉神经元在健康和再生嗅觉系统中的功能整合

• 批准号: 10630475
• 负责人: Claire Elizabeth Jane Cheetham
• 金额: $ 7.95万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-01 至 2025-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10630475
• 关键词: Adult; Area; Biological Assay; Brain; Brain Injuries; Calcium; Cell Therapy; Chemicals; Development; Electron Microscopy; Functional Imaging; Functional Regeneration; Goals; Image; Immunohistochemistry; Life; Mammals; Modeling; Natural regeneration; Neurons; Newborn Infant; Odors; Olfactory Epithelium; Olfactory Pathways; Phase; Play; Population; Research; Role; Sensory; Stem Cell Development; Stem cell transplant; Testing; Time; Work; epithelial stem cell; in vivo; innovation; insight; neural circuit; newborn neuron; olfactory sensory neurons; optogenetics; repaired; sensory input; stem cell population; stem cells; synaptogenesis; two-photon

Project Summary/Abstract Understanding how endogenously-generated adult-born neurons functionally integrate into neural circuits may inform strategies employing stem cell-derived neurons to repair damaged brain areas. Olfactory sensory neurons (OSNs), which are generated throughout life in all mammals, provide an ideal model to study functional integration in both the healthy and the regenerating olfactory system. Our long-term goal is to determine how circuit function and regeneration can be enhanced by functional integration of newborn neurons. The overall objective of this application is to determine how evoked activity during an immature developmental window influences OSN survival and integration in the healthy and the regenerating olfactory system. Our central hypothesis is that odor selective input to immature OSNs is required for their survival and functional integration in both the healthy and the regenerating olfactory system. To test this hypothesis, we propose three specific aims. First, we will define the odor selectivity of immature OSN sensory input to the OB, using in vivo 2-photon calcium imaging. Second, we will determine the role of evoked activity in immature adult-born OSNs in their survival and functional integration. We will up- and down-regulate evoked activity in OSNs during the immature developmental stage, and use immunohistochemistry, electron microscopy, optogenetics and in vivo 2-photon structural and functional imaging to assay their survival, synaptogenesis and functional integration. Finally, we will determine the role of immature OSN activity in regeneration of OSN input to the OB. For this aim, we will chemically ablate OSNs, which regenerate from olfactory epithelium stem cells, manipulate activity using naris occlusion and chemogenetic silencing during regeneration, and use in vivo 2-photon time-lapse imaging to track structural and functional regeneration of OSN input to glomeruli. We expect to find that evoked activity during immaturity plays a key role in the survival and integration of that OSN. Hence, our proposed work will have an important impact by defining how functional integration of an endogenous population of stem cell-derived neurons is regulated to maintain and regenerate brain function. This study will be highly significant in elucidating the role of activity in an endogenously-generated population of newborn neurons in regulating their functional integration, thereby providing important insight relevant to the development of stem cell-based therapeutic strategies to repair damaged neural circuits. Our proposed research is conceptually innovative in proposing that sensory input during the immature phase of development is crucial to successful functional integration of OSNs in the healthy and regenerating OB and will employ a technically innovative array of experimental approaches.

项目概要/摘要 了解内源生成的成年神经元如何功能性地整合到神经回路中可能 为利用干细胞衍生的神经元修复受损大脑区域的策略提供信息。嗅觉感觉神经元 （OSN）在所有哺乳动物的一生中都会产生，为研究功能性神经元提供了理想的模型 整合健康和再生的嗅觉系统。我们的长期目标是确定如何 新生神经元的功能整合可以增强回路功能和再生。整体 该应用程序的目的是确定在不成熟的发育窗口期间如何诱发活动 影响 OSN 在健康和再生嗅觉系统中的存活和整合。我们的中央 假设是未成熟 OSN 的气味选择性输入是其生存和功能整合所必需的 在健康和再生的嗅觉系统中。为了检验这个假设，我们提出了三个具体的 目标。首先，我们将使用体内 2 光子定义未成熟 OSN 感觉输入到 OB 的气味选择性 钙成像。其次，我们将确定诱发活动在未成熟的成年 OSN 中的作用 生存和功能整合。我们将在未成熟期间上调和下调 OSN 的诱发活动 发育阶段，并使用免疫组织化学、电子显微镜、光遗传学和体内 2-光子 结构和功能成像来测定它们的存活、突触发生和功能整合。最后，我们 将确定未成熟 OSN 活性在 OB 的 OSN 输入再生中的作用。为了这个目标，我们将 化学消融由嗅上皮干细胞再生的 OSN，利用鼻孔操纵活动 再生过程中的闭塞和化学遗传学沉默，并使用体内 2 光子延时成像来跟踪 输入肾小球的 OSN 的结构和功能再生。我们期望发现在 不成熟度对于 OSN 的生存和整合起着关键作用。因此，我们提出的工作将有一个 通过定义干细胞衍生的内源群体的功能整合如何产生重要影响 神经元受到调节以维持和再生大脑功能。这项研究对于阐明 内源性新生神经元群体的活动在调节其功能中的作用 整合，从而为基于干细胞的治疗的开发提供重要的见解 修复受损神经回路的策略。我们提出的研究在概念上具有创新性，提出 发育不成熟阶段的感觉输入对于 OSN 成功的功能集成至关重要 在健康和再生的产科中，将采用一系列技术创新的实验方法。