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

• 批准号: 8327225
• 负责人: SYLVIE BRETON
• 金额: $ 55.82万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-21 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8327225
• 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

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）：生物学中的当前范例是存在于假复层上皮中的所谓基底细胞从不与器官的腔侧接触。与这一教条相反，我们最近发现，这些细胞延伸细长的身体投影，穿过紧密连接屏障到达管腔。这项研究提案是由这一范式转变的发现驱动的。我们发现，基底细胞扫描选定的上皮细胞的腔侧，并通过与其他上皮细胞的串扰来调节它们的功能。我们观察到“顶端到达”基底细胞在几个组织中的男性生殖道和上呼吸道，表明腔采样属性的基底细胞是一个普遍的现象。在这项研究计划中，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讯网络。为了做到这一点，我们将产生新的小鼠表达红色荧光蛋白，mRaspberry，只在基底细胞。我们将这些小鼠与我们实验室现有的小鼠杂交，包括在树突细胞中表达黄色荧光蛋白的CD 11 c-YFP小鼠，以及在附睾透明细胞、肺的非纤毛细胞和肾闰细胞中表达绿色荧光蛋白EGFP的B1-EGFP小鼠。这将产生一种新的动物模型，其中几种细胞类型将同时成像，在活的动物，使用活体多光子显微镜配备了小型化的目标，并在其中离子和一氧化氮流量将被测量在真实的时间使用选择性微电极。我们将集中在两个上皮，附睾，这是我们的研究计划的核心，和气管。附睾连接睾丸和输精管，参与精子的成熟和储存，因此在男性生育力中起着至关重要的作用。本申请的另一个目标组织，气管，不断地被外来过敏性和致病性物质侵入，并在气道和身体之间提供结构屏障。我们认为，基底细胞是探测附睾中调节男性生育能力的管腔因子和气管中吸入分子的前线传感器。更好地了解基底细胞的新型顶端感觉作用以及它们如何将其发现传递到相邻细胞将有助于确定男性不育和肺部疾病（包括哮喘，慢性阻塞性肺病（COPD）和囊性纤维化（CF））的病理生理机制。因此，监测和解码附睾和气管中的细胞间对话将促进治疗这些疾病的创新诊断和治疗干预措施。此外，这项研究计划将对我们对上皮细胞的理解产生更广泛的影响，因为这里产生的数据将为复杂组织建立的通信网络以及疾病中的干扰方式提供前所未有的见解。