ENSMAP: Molecular and Functional Mapping of the Enteric Nervous System

ENMAP：肠神经系统的分子和功能图谱

• 批准号: 9531523
• 负责人: E Michelle SOUTHARD-SMITH
• 金额: $ 29.31万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-27 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9531523
• 关键词: Adult; Anatomy; Atlases; Biological Assay; Bladder; Blood Vessels; Catalogs; Cells; Cellular Morphology; Collaborations; Complement; Complex; Congenital Megacolon; Data; Data Set; Databases; Defecation; Development; Diabetic Neuropathies; Diet; Dilatation - action; Disease; Documentation; Electrodes; Electrophysiology (science); Enteral; Enteric Nervous System; Excision; Female; Fluorescence-Activated Cell Sorting; Foundations; Frequencies; Ganglia; Gastric Emptying; Gastrointestinal Motility; Gene Expression; Gene Mutation; Genes; Genetic; Goals; Health; Human; Image; Immunohistochemistry; In Situ; Inbred Strain; Inbred Strains Mice; Inflammatory disease of the intestine; Intestinal Motility; Intestines; Label; Large Intestine; Maps; Measures; Methods; Microelectrodes; Molecular; Molecular Profiling; Mouse Strains; Movement; Mus; Neuroanatomy; Neurobiology; Neurons; Nuclear; Operative Surgical Procedures; Organ; Pattern; Perfusion; Peristalsis; Pharmacologic Substance; Pharmacology; Phenotype; Physiological; Physiology; Positioning Attribute; Process; Property; Proteins; Research Personnel; Resolution; Scientist; Small Intestines; Stem cells; Surveys; System; Technology; Transgenes; Transgenic Mice; Variant; Work; base; cell motility; cell type; deep sequencing; feeding; fetal; gene complementation; human data; immunohistochemical markers; laser capture microdissection; live cell imaging; male; man; member; migration; motility disorder; mouse model; nutrient absorption; phenome; progenitor; public health relevance; relating to nervous system; response; transcriptome; transcriptome sequencing; treatment planning

PROJECT SUMMARY Gastrointestinal (GI) motility and defecation are absolute prerequisites for nutrient absorption, fecal elimination and overall health. Normal GI motility, vascular perfusion, and intestinal inflammation are coordinated by vast numbers of neurons that reside within ganglia of the enteric nervous system (ENS) intrinsic to the gut wall. While recent work has identified diverse genes that direct the initial development of progenitor cells that give rise to enteric neurons in the wall of the intestine, we know very little about the genes that are expressed in adult enteric neurons. Consequently we are unable to determine whether efforts to generate enteric neurons produce the normal complement of cell types. Moreover we do not fully understand how distinct types of neurons contribute to overall coordination of intestinal motility because the use of common immunohistochemical markers alone does not distinguish functionally distinct subtypes. As a result, our abilities to target and functionally manipulate specific types of neurons in the gut are extremely limited. To surpass these limitations, our application proposes to develop a comprehensive, single cell transcriptome map of adult enteric neurons in normal mice in parallel with deep sequencing of enteric ganglia from distinct regions of human intestine so that a global gene expression atlas of human enteric ganglia is obtained. To capture mouse enteric neurons for single cell RNA-Seq we will use a fluorescent transgenic mouse line that we developed for live-cell imaging of enteric neurons. Human enteric ganglia will be collected by laser capture microdissection from adult surgical remnants. Comparison of enteric neuron expression profiles between mouse and human data sets will identify conserved genes that mark distinct neuronal subtypes. The resulting expression atlas of enteric neurons will define specific molecular fingerprints for discrete neuron subtypes that are essential to pursue targeted, functional manipulation of GI motility in distinct regions of the intestine.

项目摘要 胃肠道（GI）运动性和排便是营养吸收，消除粪便和整体健康的绝对先决条件。 正常的胃肠道运动，血管灌注和肠炎与肠道神经神经节内（ENS）内部固有的大量神经元协调。 尽管最近的工作已经确定了指导祖细胞最初发育的多种基因，这些基因引起了肠壁中肠神经元的初步发展，但我们对成人肠神经元中表达的基因知之甚少。 因此，我们无法确定产生肠神经元的努力是否会产生细胞类型的正常补体。 此外，我们不完全理解不同类型的神经元如何促进肠道运动的总体协调，因为单独使用常见的免疫组织化学标记并不能区分功能上不同的亚型。 结果，我们靶向和操纵肠道中特定类型的神经元的能力极为有限。 为了超越这些局限性，我们的应用建议与正常小鼠中成年肠神经元的全面单细胞转录组图并行，并与人类肠道不同区域的肠神经节进行深度测序，从而获得人类肠神经神经节的全球基因表达地图。 为了捕获单细胞RNA-seq的小鼠肠神经元，我们将使用我们开发的荧光转基因小鼠系，用于肠神经元的活细胞成像。 人类肠神经节将通过激光捕获成人手术残余物的显微解剖来收集。 小鼠和人类数据集之间肠神经元表达谱的比较将识别标记不同神经元亚型的保守基因。所得的肠神经元的表达地图集将定义特定的分子指纹，用于离散的神经元亚型，这对于在肠道不同区域进行靶向的gi运动性能是必不可少的。