GROWTH AND DIFFERENTIATION OF AIRWAY GLANDS

气道腺的生长和分化

• 批准号: 6272612
• 负责人: CAROL B BASBAUM
• 金额: $ 33.79万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6272612
• 关键词: DNA binding protein; Mycoplasma; bronchial mucus; cell differentiation; cell type; cellular pathology; chemical binding; confocal scanning microscopy; disease /disorder model; gel mobility shift assay; growth /development; growth factor receptors; histogenesis; immunofluorescence technique; in situ hybridization; isozymes; laboratory rat; lysozyme; polymerase chain reaction; protein biosynthesis; respiratory epithelium; trachea; transcription factor; transforming growth factors

The airways of individuals suffering from hypersecretion exhibit gland hypertrophy as well as a serous-mucous cell shift within the glands. These reflect disease-related abnormalities in gland growth and differentiation. The goal of the proposed studies is to gain information about the mechanisms controlling these processes. So that mechanisms relevant to normal development and disease can be compared, experimental studies will be performed in two model systems (1) the trachea of the newborn rat, in which glands develop between postnatal days 7 and 28 and (2) the trachea of the mycoplasma-infected adult rat, in which new glands develop between postinfection days 7 and 42. Studies listed under Specific Aim 1 will determine by confocal microscopy the size and distribution of tracheal submucosal glands at defined postnatal and post- infection time points in vivo as well as in an in vitro gland growth explant system. Studies listed under Specific Aim 2 will identify the cell type from which gland buds arise in each model, using double immunofluorescence to define cells that are both preparing to divide and express phenotype-specific markers. They will also determine by PCR, immunofluorescence and in situ hybridization whether elevations in the local concentration of specific growth factors and/or their receptors precede mitosis in these cells. Evidence for causal relationships between specific growth factors and gland growth will be sought using neutralizing antibodies in the in vitro gland growth explant system. Studies listed under Specific Aim 3 will identify extracellular matrix- degrading enzymes whose local tissue concentrations are elevated at the time of gland growth in each model and seek evidence for causal relationships between specific enzymes and gland growth using neutralizing antibodies in the in vitro gland growth explant system. Mechanisms controlling enzymes relevant to gland growth will be identified. Studies listed under Specific Aim 4 will examine serous differentiation mechanisms by identifying DNA-protein interactions controlling serous cell-specific lysozyme transcription. Rat lysozyme cDNAs and genomic clones will be isolated, the transcription start site mapped, and regulatory DNA sequences identified using routine methods. DNA regulatory sequences specifically involved in the initiation of lysozyme transcription during serous cell differentiation will be identified using a novel method of in vivo footprinting. Transcription factors interacting with these sequences will be isolated. Knowledge gained from these studies will hopefully suggest pharmacological interventions to inhibit excessive gland growth and differentiation abnormalities associated with hypersecretory airways.

遭受超分分泌的个体的呼吸道表现出腺体 肥胖以及腺体内的浆液性细胞移位。 这些反映了与疾病相关的异常，腺体生长和 分化。 拟议研究的目的是获取信息 关于控制这些过程的机制。 这样的机制 可以比较与正常发育和疾病有关的实验 研究将在两个模型系统中进行（1） 新生大鼠，腺体在产后第7和28天之间发育 （2）成年大鼠的支原体感染的成年大鼠的气管 在感染后第7和第42天发展。 特定的目标1将通过共聚焦显微镜大小和 在定义的产后和 - 感染时间点体内以及体外腺体生长 外植体系统。 在特定目标2下列出的研究将确定 使用双 免疫荧光以定义准备分裂的细胞和 特定表型特异性标记。 他们还将通过PCR确定 免疫荧光和原位杂交是否升高 特定生长因子和/或其受体的局部浓度 在这些细胞中有丝分裂之前。 因果关系的证据 在特定的生长因素和腺体之间的生长之间将寻求 中和体外腺体生长外植体系统中的抗体。 在特定目标3下列出的研究将确定细胞外基质 - 降解酶的局部组织浓度在 每个模型的腺体生长时间，并寻求因果关系的证据 使用特定酶与腺体生长之间的关系 中和体外腺体生长外植体系统中的抗体。 控制与腺体生长有关的酶的机制将是 确定。 在特定目标4下列出的研究将检查浆液 通过识别DNA-蛋白质相互作用的分化机制 控制浆液细胞特异性溶菌酶转录。 大鼠溶菌酶 将分离cDNA和基因组克隆，转录起始位点 映射，使用常规方法鉴定的调节DNA序列。 DNA调节序列特别涉及 浆液细胞分化过程中的溶菌酶转录将是 使用一种新型的体内足迹方法鉴定。 转录 与这些序列相互作用的因素将被隔离。 知识 从这些研究中获得的有望提出药理学 抑制过度腺体生长和分化的干预措施 异常与超偏呼吸道有关。