Linking the molecular logic of sensory neuron diversity and somatosensory circuitry in mouse spinal cord: development of novel tools for viral tracing and 3D analysis

将小鼠脊髓感觉神经元多样性的分子逻辑与体感回路联系起来：开发病毒追踪和 3D 分析的新工具

• 批准号: 10307604
• 负责人: JANE DODD
• 金额: $ 20.25万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 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 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10307604
• 关键词: 3-Dimensional; 3D Print; Adult; Afferent Neurons; Anatomy; Atlases; Axon; Behavior; Brain; Cells; Computer software; Custom; Data; Development; Disease; Dorsal; Embryo; Event; Gene Expression; Gene Expression Profile; Genetic; Growth; Health; Helper Viruses; Histologic; Histology; Image; Immunohistochemistry; In Situ; Individual; Injections; Injury; Interneurons; Intervention; Limb structure; Link; Logic; Maps; Methods; Modality; Molecular; Molecular Analysis; Molecular Profiling; Mus; Musculoskeletal Equilibrium; Nervous system structure; Neurons; Pathway interactions; Population; Positioning Attribute; Procedures; Proteins; Rabies virus; Resolution; Resources; Sensory; Site; Spatial Distribution; Spinal; Spinal Cord; Spinal Ganglia; Synapses; System; Three-dimensional analysis; Time; Tracer; Transgenic Organisms; Viral; Virus; Water; blastomere structure; computerized tools; design; experimental study; in utero; insight; mature animal; molecular array; novel; postnatal; programs; promoter; retrograde transport; single-cell RNA sequencing; somatosensory; tau Proteins; tool; trait; uptake

Project Summary/Abstract Sensory neurons in the dorsal spinal cord (dINs) represent the first site in the CNS to receive somatosensory information from the periphery and are a crucial component in sensorimotor circuits that control posture and behavior. Defining the developmental principles of somatosensory circuit formation and the molecular identity of neurons that serve specific sensory functions is key to our understanding of the logic of sensorimotor integration and for strategic interventions following injury or disease. All dINs arise from a small array of molecularly defined embryonic cell populations, but markers are expressed only transiently and how these early neurons diversify and integrate into sensory circuits serving distinct sensory modalities in the mature animal remains largely unknown. There is an urgent need for genetic access to express tracers and to manipulate individual classes of developing dINs as they emerge during the embryonic period but establish circuits postnatally. Here, we will generate for the first time, mice that bridge the gap between early identity and mature function by providing access selectively to one population of dINs, called dI1s, throughout development. We will create mouse lines to introduce anterograde and retrograde viral tracers to reveal and map the synaptic targets of dI1s in brain and spinal cord, and to identify input neurons to dI1s, in DRG, brain and spinal cord. These lines will extend temporal genetic access selectively to developing dI1s into postnatal stages, by, first, a Cre-dependent Cre approach, permitting access by Cre-dependent axonal and synaptic tracers, and second, mice that express the proteins required for transsynaptic tracing using rabies virus. We will also develop platforms to analyze efficiently the anatomy and connectivity of the circuitry. Furthermore, we will create a widely needed 3D spinal cord atlas for assessment of neuronal identity and mapping circuitry. To understand the programs that regulate dI1 development, we will also identify and follow dynamic changes of transcriptional signatures in dI1s throughout embryonic and postnatal ontogeny by single cell RNA sequencing. The tools established in the proposed experiments and analyses will be applicable to all dIN subsets and will contribute both resources and information to the field, and provide a template for manipulation and functional analysis of somatosensory pathways in health and in disease.

项目摘要/摘要 背脊髓（DIN）中的感觉神经元代表CNS中的第一个接收体感 来自外围的信息，是控制姿势和 行为。定义体感电路形成的发展原理和分子身份 具有特定感觉功能的神经元是我们对感觉运动整合逻辑的理解的关键 以及受伤或疾病后的战略干预措施。所有DIN都来自一小部分分子定义 胚胎细胞种群，但仅暂时表达标记，这些早期神经元如何多样化 并集成到成熟动物中具有独特感官方式的感觉电路中很大程度上保持 未知。迫切需要遗传获取来表达示踪剂并操纵单个类别 在胚胎时期出现时出现DIN，但在产后建立电路。在这里，我们会的 首次生成的小鼠通过提供弥合早期身份和成熟功能之间的差距 在整个开发过程中，有选择地访问一个名为DI1的DIN人群。我们将创建鼠标线 引入顺行和逆行病毒示踪剂，以揭示和映射大脑中DI1的突触靶标 脊髓，并在DRG，脑和脊髓中鉴定DI1的输入神经元。这些线将延长暂时性 遗传访问选择性地将DI1分为产后阶段，首先是CRE依赖性CRE方法， 允许通过CRE依赖性轴突和突触示踪剂进入，其次是表达蛋白质的小鼠 使用狂犬病病毒进行跨色球跟踪所必需的。我们还将开发平台以有效分析 解剖学和电路的连通性。此外，我们将创建一个广泛需要的3D脊髓图集 评估神经元身份和映射电路。了解规范DI1的程序 开发，我们还将确定并遵循DI1中转录特征的动态变化 单细胞RNA测序的胚胎和产后个体发育。提议中建立的工具 实验和分析将适用于所有DIN子集，并将贡献资源和信息 到达现场，并提供一个模板，以操纵和功能分析健康中的体感途径 和疾病。