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：
8692557
负责人：
KENTON M SANDERS
金额：
$ 41.72万
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 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8692557
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 public health relevance purine relating to nervous system response

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（GI）肌肉中的神经肌肉装置比以前理解的更复杂。肠运动神经末梢与 3 种细胞密切接触：平滑肌细胞 (SMC)、卡哈尔间质细胞 (ICC) 和成纤维细胞样细胞 (FLC)。 ICC 和FLC 均与SMC 电耦合。因此，由 ICC 和 FLC 转导的信号可以在整个 SMC 合胞体中进行电传递。过去有大量研究针对 SMC 和 ICC，但很少有研究探讨 FLC 在胃肠道器官中的功能。该项目将评估 FLC 在小鼠和人类胃肠道肌肉中的功能。 FLC 专门标记有 PDGFR-α 抗体。我们开发并验证了在 PDGFR-α+ 细胞中持续表达 eGFP 的转基因小鼠模型。我们将分离 PDGFR-α+ 细胞，并使用膜片钳技术来研究这些细胞表达的内源性电导，并表征肠道神经递质对膜电导的影响。初步研究表明，嘌呤神经递质会激活大量外向电流，并将研究该电流的特性和药理学。 SMC 和 ICC 通过激活净内向电流来响应嘌呤递质。因此，我们的数据表明 PDGFR-α 细胞介导胃肠道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 + 细胞的神经支配。我们还将使用荧光激活细胞分选来纯化 PDGFR-α+ 细胞并表征 PDGFR+ 细胞的转录组。还将研究失去 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神经调节的理解，并可能为结肠运动和功能性胃肠道治疗提供新的靶点失调。