BRONCHIAL SECRETIONS: PHYSICAL & CHEMICAL STUDIES

支气管分泌物：物理

• 批准号: 3344757
• 负责人: Mary C Rose
• 金额: $ 21.36万
• 依托单位: CHILDREN'S NATIONAL MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-03-01 至 1997-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3344757
• 关键词: bronchial mucus; cell growth regulation; gene expression; genetic library; genetic mapping; genetic regulatory element; genetic transcription; glycoproteins; glycosyltransferase; human tissue; immunologic assay /test; in situ hybridization; laboratory rabbit; lung disorder; molecular cloning; molecular pathology; mucins; northern blottings; nucleic acid sequence; protein sequence; protein structure function; respiratory epithelium; tissue /cell culture

Mucin glycoproteins (mucins), the major components of the mucosal layer that protects and lubricates mammalian airways, are secreted by epithelial cells lining the lumen and submucosal glands. These cells and glands exhibit hyperplasia in chronic obstructive pulmonary diseases. Little is known about the mechanism or factors initiating or regulating metaplasia and hyperplasia, although it has recently been proposed that initiation of transcription of mucin protein genes by serous cells in rat airways is a primary event leading to goblet and mucous cell metaplasia and hypersecretion (1). Current evidence indicates that several mucin genes are expressed both in the airways and intestine. We speculate that the specific mucin proteins are more highly expressed in airway diseases (perhaps by specific airway cells) and that different members of the airway mucin protein gene family provide unique substrates for specific glycosyltransferases. The long term objective of our research program is to elucidate the role of airway mucins in health and in diseases. We have recently identified and characterized a cDNA clone that encodes TBM, a human tracheobronchial mucin that appears to be airway-specific. The specific aims of this proposal are: [1] To identify and characterize full length cDNA clones encoding TBM and to delineate similarities and differences to other mucins as such information becomes available. [2] To elucidate the cell and tissue specificity of mucin genes by in situ hybridization and Northern blot analysis and determine whether the expression of specific mucin genes is affected in airway diseases. [3] To isolate and characterize genomic DNA for TBM. The cDNA clone that encodes for TBM will be used to probe genomic libraries. TBM genomic clones will be characterized by chromosomal location, restriction mapping, S1 mapping, and by sequence analysis of the exon/intron boundaries. [4] To investigate expression and regulation of the TBM gene. We will determine the transcriptional activity of the TBM gene. Should it prove transcriptionally active, the promoter sequences of the genomic clones identified in aim [3] will be studied for regulatory elements using promoter deletion constructs.

粘蛋白糖蛋白（粘蛋白），粘膜层的主要成分 保护和润滑哺乳动物呼吸道，由 管腔和粘膜下腺内衬的上皮细胞。 这些细胞 慢性阻塞性肺疾病中腺体增生 疾病。 对于引发或发生的机制或因素知之甚少 调节化生和增生，尽管最近它已被 提出粘蛋白基因转录的起始 大鼠气道中的浆液细胞是导致杯状和 粘液细胞化生和分泌过多 (1)。 目前的证据 表明一些粘蛋白基因在气道和气道中都有表达 肠。 我们推测特定的粘蛋白更多 在气道疾病中高表达（可能由特定气道细胞表达） 气道粘蛋白基因家族的不同成员 为特定的糖基转移酶提供独特的底物。 我们研究计划的长期目标是阐明其作用 气道粘蛋白在健康和疾病中的作用。 我们最近确定了 并鉴定了编码 TBM 的 cDNA 克隆，TBM 是一种人类 气管支气管粘蛋白似乎具有气道特异性。 具体的 该提案的目标是： [1] 识别并表征全长 编码 TBM 的 cDNA 克隆并描述相似点和差异 当此类信息可用时，可将其用于其他粘蛋白。 [2] 阐明 通过原位杂交研究粘蛋白基因的细胞和组织特异性 和 Northern blot 分析并确定是否表达 特定的粘蛋白基因在气道疾病中受到影响。 [3] 隔离和 表征 TBM 的基因组 DNA。 编码 TBM 的 cDNA 克隆 将用于探测基因组文库。 TBM 基因组克隆将 以染色体位置、限制性图谱、S1图谱为特征， 以及外显子/内含子边界的序列分析。 [4] 至 研究 TBM 基因的表达和调控。 我们将 确定 TBM 基因的转录活性。 是否应该证明 转录活性，基因组克隆的启动子序列 将使用目标 [3] 中确定的监管要素来研究 启动子缺失构建体。