PLASMA MEMBRANE REGIONS OF CELLS GROWN AS MONOLAYERS

单层生长细胞的质膜区域

• 批准号: 2175400
• 负责人: BRUCE Shell JACOBSON
• 金额: $ 15.37万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-04-01 至 1997-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2175400
• 关键词: HeLa cells; antibody; arachidonate; cell adhesion; cell membrane; cell migration; collagen; eicosanoid metabolism; high performance liquid chromatography; integrins; laboratory mouse; lipoxygenase; neoplastic cell culture for noncancer research; nucleic acid sequence; phospholipids; phosphorylation; protein kinase C; protein sequence; protein structure function; receptor; recombinant DNA; second messengers; transfection; HeLa cells; antibody; arachidonate; cell adhesion; cell membrane; cell migration; collagen; eicosanoid metabolism; high performance liquid chromatography; integrins; laboratory mouse; lipoxygenase; neoplastic cell culture for noncancer research; nucleic acid sequence; phospholipids; phosphorylation; protein kinase C; protein sequence; protein structure function; receptor; recombinant DNA; second messengers; transfection

The long term goal of this project has been to determine the sequence and regulation of the molecular events that take place when cells attach and spread on extracellular matrices to form two distinct plasma membrane domains; one domain is attached to the matrix and the other is exposed to the extracellular milieu. Attachment and spreading of cells is involved in a number of biological phenomena including embryogenesis, blood coagulation, wound healing, and metastasis. We have been using HeLa cell attachment and spreading on collagen as a model to study this problem. Results obtained during the last funding period have led us to hypothesize the following: Cell-substrate attachment induces the exocytotic upregulation of collagen receptors. Concurrently, substrate induced clustering of three non-integrin type collagen receptors as well as B1 integrin collagen receptors drives the receptors to bind to the cytoskeleton. Receptor clustering also turns on a sequence of several second messengers and feedback mechanisms to initiate and optimize cell spreading. Our current objectives are to: (1) Determine whether the three HeLa cell non-integrin collagen receptors are a family of receptors different than the integrins. (2) Characterize the pathways responsible for producing the sequence of second messengers that initiate HeLa cell spreading on a gelatin substrate. HeLa cells exhibit three variables during cell spreading; rate of spreading, percentage of cells that spread, and the extent of spreading. The goal is not only to fully characterize the basic second messenger sequence regulating cell spreading but to also begin to determine if there are subsets of second messengers that could regulate the different variables of spreading. (3) Determine whether the three non-integrin and the B1 integrin collagen receptors in HeLa cells act independently, additively, or synergistically on the same or different second messengers to facilitate HeLa cell spreading. Lastly, we will begin work to determine if what we observe with the regulation of HeLa cell spreading, also occurs with more complex cells such endothelial cells.

该项目的长期目标是确定顺序和 调节细胞附着时发生的分子事件和 在细胞外矩阵上散布以形成两个不同的质膜 域；一个域附着在矩阵上，另一个域暴露于 细胞外环境。细胞的附着和扩散与 许多生物学现象，包括胚胎，血液 凝结，伤口愈合和转移。 我们一直在使用HeLa细胞 附着并在胶原蛋白上扩散，作为研究此问题的模型。 在上一个资金期间获得的结果使我们假设 以下内容：细胞基底附着诱导胞吐 胶原受体的上调。 同时诱导底物 三种非整合蛋白型胶原蛋白受体的聚类以及B1 整合素胶原受体驱动受体与 细胞骨架。受体聚类也打开了几个序列 启动和优化单元格的第二个使者和反馈机制 蔓延。我们当前的目标是：（1）确定三个目标是否 HeLa细胞非整合蛋白胶原蛋白受体是一个受体家族 与整合素不同。 （2）表征负责的途径 用于产生启动HeLa细胞的第二个使者的顺序 扩展在明胶底物上。 HeLa细胞表现出三个变量 在细胞扩散期间；扩散率，传播的细胞百分比， 以及扩散的程度。目标不仅是充分表征 基本的第二信使序列调节细胞扩散，但也 开始确定是否有第二个使者的子集可以 调节不同的扩散变量。 （3）确定是否 HeLa细胞中的三个非整合蛋白和B1整合素胶原受体 在相同或不同的相同或不同 第二使者促进Hela细胞扩散。 最后，我们会的 开始工作以确定我们对HeLa的调节是否观察到的 细胞扩散，也发生了更复杂的细胞，例如内皮 细胞。