3-Dimensional modeling of basal cell function in pseudostratified epithelia

假复层上皮基底细胞功能的三维建模

基本信息

批准号：
8541004
负责人：
SYLVIE BRETON
金额：
$ 53.2万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-21 至 2015-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8541004
关键词：
3-Dimensional AQP9 gene Air Animal Model Animals Apical Asthma Basal Cell Behavior Biological Models Biology Blood Breathing Breeding Cell Line Cell physiology Cells Characteristics Chemicals Chronic Obstructive Airway Disease Clear Cell Color Communication Complex Crossbreeding Cyan Fluorescent Protein Cystic Fibrosis Data Dendritic Cells Development Diagnostic Disease Electrodes Epididymis Epithelial Cells Epithelium Female Fertility Future Genes Gland Goals Green Fluorescent Proteins Hormones Human ITGAX gene Image Individual Intercalated Cell Invaded Ion-Selective Electrodes Ions Kidney Knowledge Laboratories Larynx Laser Scanning Confocal Microscopy Life Liquid substance Lung Lung diseases Male Infertility Measures Microelectrodes Microscope Modeling Monitor Mus Nitric Oxide Olfactory Mucosa Organ Organism Pharmaceutical Preparations Play Production Property Prostate Proteins Pseudostratified Epithelium Research Research Proposals Role Sampling Scanning Sensory Side Stimulus Structure System Systems Biology Testis Therapeutic Intervention Three-Dimensional Imaging Tight Junctions Time Tissue Model Tissues Trachea Transgenic Mice Tube United States National Institutes of Health Upper respiratory tract Vas deferens structure Vision cell type in vivo innovation insight intercellular communication male miniaturize mouse model novel novel diagnostics novel strategies novel therapeutics pathogen programs promoter red fluorescent protein reproductive response sensor sperm cell time use tool water channel

项目摘要

DESCRIPTION (provided by applicant): A current paradigm in biology is that so-called basal cells, present in pseudostratified epithelia, are never in contact with the luminal side of an organ. In contrast to this dogma, we recently showed that these cells extend slender body projections that cross the tight-junction barrier to reach the lumen. This research proposal is driven by this paradigm-shifting discovery. We found that basal cells scan the luminal side of selected epithelia and modulate their function via crosstalk with other epithelial cells. We observed "apical-reaching" basal cells in several tissues of the male reproductive and upper respiratory tracts indicating that the luminal sampling property of basal cells is a generalized phenomenon. In this research program, we will examine the spatial, temporal and motional behavior of basal cells in vivo and we will characterize the intercellular communication networks between basal cells and adjacent cells. To do so, we will generate novel mice expressing the red fluorescent protein, mRaspberry, in basal cells exclusively. We will cross-breed these mice with current mice available in our laboratory, including CD11c-YFP mice that express the yellow fluorescent protein in dendritic cells, and B1-EGFP mice that express the green fluorescent protein EGFP in epididymal clear cells, non- ciliated cells of the lung, and kidney intercalated cells. This will generate a novel animal model in which several cell types will be imaged simultaneously, in live animals, using intravital multiphoton microscopes equipped with miniaturized objectives, and in which ionic and nitric oxide fluxes will be measured in real time using selective microelectrodes. We will focus on two epithelia, the epididymis, which is at the core of our research program, and the trachea. The epididymis, which connects the testis to the vas deferens, is involved in the maturation and storage of spermatozoa and, therefore, plays a crucial role in male fertility. The other target tissue of this application, the trachea, is constantly invaded by foreign allergenic and pathogenic substances, and provides a structural barrier between the airway and the body. We propose that basal cells are front-line sensors that probe luminal factors that regulate male fertility in the epididymis, and inhaled molecules in the trachea. A better understanding of the novel apical sensory role of basal cells and how they transmit their findings to adjacent cells will help define the pathophysiological mechanisms underlying male infertility, and diseases of the lung, including asthma, chronic obstructive pulmonary disease (COPD) and cystic fibrosis (CF). Monitoring and decoding intercellular conversations in the epididymis and trachea will, thus, promote innovative diagnostic and therapeutic interventions for the treatment of these diseases. In addition, this research program will have broader implications for our understanding of epithelia in general, as data generated here will provide unprecedented insights into the communication network established by complex tissues and on how it is perturbed in disease.

描述（由申请人提供）：生物学的当前范例是，存在于假性上皮中的所谓基底细胞从未与器官的腔侧接触。与这种教条相反，我们最近表明这些细胞扩展了细长的身体投影，这些投影穿过紧密结的屏障以达到管腔。该研究建议是由这种范式转移发现的驱动的。我们发现基底细胞扫描了选定上皮的腔侧，并通过与其他上皮细胞串扰调节其功能。我们观察到男性生殖和上呼吸道的几个组织中的“根尖”基底细胞，表明基底细胞的腔采样特性是一种普遍的现象。在该研究计划中，我们将研究体内基底细胞的空间，时间和运动行为，我们将表征基底细胞和相邻细胞之间的细胞间通信网络。为此，我们将仅在基底细胞中生成表达红色荧光蛋白Mraspberry的新小鼠。我们将与实验室中可用的电流小鼠交叉杂交，包括在树突状细胞中表达黄色荧光蛋白的CD11C-YFP小鼠，以及在附子清除细胞中表达绿色荧光蛋白EGFP的B1-EGFP小鼠。这将产生一种新型的动物模型，其中将在活动物中同时使用含有微型目标的浸润性多光子显微镜同时成像，并使用选择性微电极实时测量离子和一氧化物通量。我们将重点关注两个上皮的附加膜，这是我们研究计划的核心和气管的核心。将睾丸连接到VAS延迟的附睾参与精子的成熟和储存，因此在男性生育中起着至关重要的作用。该应用的另一个目标组织，气管，不断被外国过敏和致病物质侵入，并在气道和人体之间提供结构性屏障。我们建议基底细胞是前线传感器，可探测腔内因子的腔内因子，这些因子调节了附睾中男性的生育能力，并且在气管中吸入了分子。更好地理解基础细胞的新型根尖感官作用以及它们如何将其发现传输到相邻细胞将有助于定义男性不育症的病理生理机制，以及肺部的疾病，包括哮喘，哮喘，慢性阻塞性肺疾病（COPD）和囊肿纤维化（CF）。因此，在附睾和气管中监测和解码细胞间对话将促进创新的诊断和治疗性干预措施，以治疗这些疾病。此外，该研究计划将对我们对上皮的理解具有更广泛的影响，因为此处生成的数据将提供对复杂组织建立的通信网络的前所未有的见解以及对疾病的扰动。