STRUCTURE AND FUNCTION OF NON BASEMENT MEMBRANE LAMININS

非基底膜层粘连蛋白的结构和功能

• 批准号: 6351902
• 负责人: ROBERT E BURGESON
• 金额: $ 39.01万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-01 至 2002-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6351902
• 关键词: Macaca mulatta; animal tissue; baboons; cow; developmental neurobiology; genetically modified animals; integrins; laboratory mouse; laboratory rat; laminin; mucopolysaccharides; protein structure function; proteoglycan; tissue /cell culture

A novel laminin subunit chain, gamma3, has been recently identified. Biochemical studies of gamma3 containing laminins show that gamma3 can associate with at least alpha2, alpha4, beta1, and beta2 subunit chains. The tissue distribution of gamma3 shows it to be present in regions not containing ultrastructurally defined basement membranes. These regions include the brain matrix and the apical surface of ciliated epithelial cells. The following specific aims describe proposed intentions to test the hypothesis that gamma3 containing laminins form a unique matrix at these non-basement membrane sites, the function of which is to stabilize the cytoskeletal arrangements required for the assembly and stabilization of cilia, and to also provide a distensible matrix stabilizing the structure of the CNS. To test this hypothesis specific aims are proposed: (1) Carefully describe the anatomical distribution of gamma3 and other selected laminin chains, as well as of known matrix macromolecules and receptors in the brain and at ciliated epithelial surfaces during development and in the adult mouse; (2) Determine the molecular composition of gamma3 containing laminins in the brain and at ciliated epithelial surfaces. Chemical quantities of the appropriate laminins will be produced in cell culture and purified. (3) Potential interactions between molecules which colocalized spatially and temporally will be tested in vitro. Binding to potential receptors will be determined by cell binding studies. (4) A gamma3 null mouse will be generated by homologous recombination and the phenotype characterized. We will cross the gamma3 and beta2 heterozygous mice to produce the double KO. It is likely that these studies will provide the foundations for further study of a novel extracellular matrix. It is also possible that these studies will provide novel insights into the pathophysiology of neurodegenerative diseases and infertility.

最近已经确定了一种新型的层粘连蛋白亚基链Gamma3。 生化 含层粘连蛋白的γ3的研究表明，γ3可以与AT相关联 至少α2，alpha4，beta1和beta2亚基链。 γ3的组织分布 表明它存在于不包含超大结构的区域中 膜。这些区域包括大脑基质和 纤毛上皮细胞。以下具体目的描述了提议 测试含有粘液蛋白蛋白的γ3的假设的意图是独特的 这些非基础膜位点的矩阵，其功能是 稳定组件所需的细胞骨架布置 纤毛的稳定，还提供可扩展的基质稳定 中枢神经系统的结构。为了检验该假设的特定目的： （1）仔细描述γ3和其他选定的解剖分布 层粘连蛋白链，以及已知的基质大分子和受体 发育期间和成年人的大脑和纤毛上皮表面 老鼠; （2）确定在 大脑和纤毛上皮表面。适当的化学量 层粘连蛋白将在细胞培养中产生并纯化。 （3）潜在的相互作用 将在空间和时间上共定位的分子之间进行测试 体外。与潜在受体的结合将由细胞结合研究确定。 （4）将通过同源重组和 表型的特征。我们将越过gamma3和beta2杂合子 小鼠产生双KO。这些研究可能会提供 进一步研究新型细胞外基质的基础。也可能 这些研究将提供对有关的新见解 神经退行性疾病和不育。