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）运动和排便是营养吸收，粪便排泄和整体健康的绝对先决条件。 正常的胃肠道运动、血管灌注和肠道炎症由位于肠壁固有的肠神经系统（ENS）神经节内的大量神经元协调。 虽然最近的工作已经确定了指导肠壁中产生肠神经元的祖细胞的初始发育的不同基因，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产生正常的细胞类型的补充。 此外，我们并不完全了解不同类型的神经元如何促进肠道运动的整体协调，因为单独使用常见的免疫组织化学标记物并不能区分功能不同的亚型。 因此，我们靶向和功能性操纵肠道中特定类型神经元的能力非常有限。 为了超越这些限制，我们的申请提出开发正常小鼠中成年肠神经元的全面的单细胞转录组图谱，同时对来自人肠的不同区域的肠神经节进行深度测序，从而获得人肠神经节的全局基因表达图谱。 为了捕获用于单细胞RNA-Seq的小鼠肠神经元，我们将使用我们开发用于肠神经元的活细胞成像的荧光转基因小鼠系。 将通过激光捕获显微切割从成人手术残留物中收集人肠神经节。 比较小鼠和人类数据集之间的肠神经元表达谱将鉴定标记不同神经元亚型的保守基因。由此产生的肠神经元表达图谱将定义离散神经元亚型的特定分子指纹，这些神经元亚型对于在肠的不同区域中追求GI运动的靶向功能操纵是必不可少的。