Functional role of fibroblast-like cells in GI muscles

胃肠道肌肉中成纤维细胞样细胞的功能作用

基本信息

批准号：
8833275
负责人：
KENTON M SANDERS
金额：
$ 40.67万
依托单位：
UNIVERSITY OF NEVADA RENO
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-01 至 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8833275
关键词：
Adverse effects Alpha Cell Antibodies Biology Cell Separation Cell physiology Cells Characteristics Colon Coupled Data Disease Dyes Enteral Enteric Nervous System Fibroblasts Fluorescence Fluorescence-Activated Cell Sorting Functional Gastrointestinal Disorders Functional disorder Gastrointestinal Motility Gastrointestinal tract structure Gene Expression Profile Giant Cells Health Human Image In Situ Inflammation Interstitial Cell of Cajal Ion Channel Label Mediating Membrane Motor Motor Neurons Movement Mus Muscle Myenteric Plexus Myopathy Nature Nerve Neurons Neurotransmitters Organ Outcome PDGFRB gene Patch-Clamp Techniques Pattern Pharmacology Phase Phenotype Physiological Physiology Plague Population Problem Solving Purines Regulation Role Signal Pathway Signal Transduction Smooth Muscle Smooth Muscle Myocytes Stretching Techniques TimeLine Transgenic Mice Work base cell motility cell type effective therapy gastrointestinal improved interstitial cell macrophage molecular phenotype motility disorder mouse model nerve supply neuromuscular neuroregulation neurotransmission novel purine relating to nervous system response targeted treatment

项目摘要

DESCRIPTION (provided by applicant): The neuromuscular apparatus in gastrointestinal (GI) muscles is more complicated than previously understood. Enteric motor nerve terminals make very close contacts with 3 types of cells: smooth muscle cells (SMC), interstitial cells of Cajal (ICC) and flibroblast-like cells (FLC). Both ICC and FLC are electrically coupled to SMC. Thus, signals transduced by ICC and FLC can be conveyed electrically throughout the syncytium of SMC. Considerable study has been directed toward SMC and ICC in the past, but there are few studies that have investigated the function of FLC in GI organs. This project will evaluate the function of FLC in mouse and human GI muscles. FLC are specifically labeled with antibodies for PDGFR-alpha. We have developed and verified a transgenic mouse model with constitutive expression of eGFP in PDGFR-alpha+ cells. We will isolate the PDGFR-alpha+ cells and use patch clamp techniques to study the endogenous conductances expressed by these cells and characterize the effects of enteric neurotransmitters on membrane conductances. Preliminary studies demonstrate a large outward current activated by purine neurotransmitters, and the characteristics and pharmacology of this current will be investigated. SMC and ICC respond to purine transmitters with activation of net inward current. Thus, our data suggest that PDGFR-alpha cells mediate purinergic neural responses in GI muscles. We will investigate the innervation of PDGFR + cells using the fluorescence of Ca2+ dyes. We will also use fluorescence activated cell sorting to purify PDGFR-alpha+ cells and characterize the transcriptome of PDGFR + cells. The effects of losing PDGFR + cells on purinergic responses will also be investigated. These studies will improve our understanding of neural regulation of colonic motility and will possibly provide new targets for therapies for motility and functional GI disorders.

描述（由申请人提供）：胃肠道（GI）肌肉中的神经肌肉设备比以前了解的要复杂。肠运动神经末端与3种类型的细胞非常紧密接触：平滑肌细胞（SMC），Cajal（ICC）的间质细胞（ICC）和氟布斯特样细胞（FLC）。 ICC和FLC均电与SMC电气耦合。因此，ICC和FLC传递的信号可以在整个SMC的合成中进行电传输。过去，大量研究针对SMC和ICC，但是很少有研究研究了FLC在GI器官中的功能。该项目将评估FLC在小鼠和人类GI肌肉中的功能。 FLC用PDGFR-Alpha的抗体特异性标记。我们已经开发并验证了一种具有EGFP在PDGFR-Alpha+细胞中的本构表达的转基因小鼠模型。我们将分离PDGFR-Alpha+细胞，并使用斑块夹技术研究这些细胞表达的内源性电导，并表征肠神经递质对膜电导的影响。初步研究表明，嘌呤神经递质激活的大型外部电流将研究该电流的特征和药理学。 SMC和ICC用净向内电流激活嘌呤发射器。因此，我们的数据表明，PDGFR-α细胞介导GI肌肉中的嘌呤能神经反应。我们将使用Ca2 +染料的荧光研究PDGFR +细胞的神经。我们还将使用荧光激活的细胞分选纯度净化PDGFR-Alpha +细胞并表征PDGFR +细胞的转录组。也将研究失去PDGFR +细胞对嘌呤能反应的影响。这些研究将提高我们对结肠运动的神经调节的理解，并可能为运动性和功能性GI提供新的靶标。疾病。