Core--Cytoskeletal Facility

核心--细胞骨架设施

• 批准号: 6660107
• 负责人: ANIRBAN BANERJEE
• 金额: $ 15.83万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6660107
• 关键词: 3T3 cells; G protein coupled receptor kinase; biological signal transduction; biomedical facility; clathrin; confocal scanning microscopy; cytoskeleton; digital imaging; enzyme induction /repression; fluorescence resonance energy transfer; human tissue; immunocytochemistry; macrophage; mitogen activated protein kinase; multiple organ failure; phosphorylation; protein kinase C; protein structure function; receptor mediated endocytosis; tissue /cell culture; trauma

Trauma provokes the release of a large number of bioactive mediators, such as lipids, hormones and cytokines (Project I-A). These ligands elicit immediate changes in function and metabolism, but also modify the cellular response(s) to subsequent stimuli (both destructively or constructively). The mechanism(s) of these adapted responses (termed priming or preconditioning, respectively) remain the focus of our investigations (with Projects II, III, IV, VII, and IX). Deservedly, ligand activated signaling by second messengers, ion-flux and protein phosphorylation continue to receive extensive study. But ligand binding also triggers receptor internalization. Using transformed models we (and others) now accept that activated receptors for several classes of bioactive mediators (such as catechols, lipids and TNFa) are internalized by clathrin coated vesicles. Clathrin coated endosomes are transported about the cell on microtubular tracks. Surprisingly, recent studies indicate that receptor internalization is necessary to initiate a novel phase of signaling. Signals emanating from internalized G-protein coupled receptors (GPCR) affect transcriptionally directed protein kinase cascades (including PKC and the MAPK modules) causing sustained changes in cell responsiveness (with Project V). However it is unclear how previous signaling events influence the interactions between internalized receptors, the endocytotic machinery and the microtubular cytoskeleton, in mammalian, especially human cells. Moreover it is unclear whether pro-inflammatory cells such as macrophages respond similarly to vulnerable targets such as endothelial or smooth muscle cells (with Project VI). We hypothesize that circulating mediators (adrenergic, lysolipid and cytokines ) (with Projects V, VII and IX) released after trauma augment internalization for membranes, receptors and subsequent signal traffic. We initially focus on ligands selected for i. pertinence to trauma, ii. mediation by GPCR, and iii, activation of PKC and MAPK modules. We will examine how these test ligands influence a. constitutive membrane/receptor trafficking and b. internalization dependent signaling in a set of myeloid and non myeloid cells from humans and rodents. These studies will form the basis for evaluating whether currently accessible therapeutic strategies can be directed against clathrin coated endocytosis and microtubule based trafficking.

创伤引起了大量生物活性介质的释放，例如脂质，激素和细胞因子（I-A项目）。这些配体会引起功能和代谢的立即变化，但也会改变细胞反应对随后的刺激（毁灭性或建设性）。这些改编的反应的机制（分别称为启动或预处理）仍然是我们研究的重点（与项目II，III，IV，IV，VII和IX相关）。值得一提的是，第二信使，离子 - 液和蛋白质磷酸化的激活信号传导继续接受广泛的研究。但是配体结合也触发受体内在化。使用转化的模型，我们（和其他）现在接受了几类生物活性介质（例如儿茶酚，脂质和TNFA）的激活受体被网状涂层囊泡内化。网格蛋白涂层的内体在微管轨道上在细胞周围运输。令人惊讶的是，最近的研究表明，受体内在化对于启动新型信号阶段是必要的。来自内部G蛋白偶联受体（GPCR）发出的信号会影响转录定向的蛋白激酶级联反应（包括PKC和MAPK模块），从而导致细胞反应性持续变化（Project V）。但是，尚不清楚以前的信号事件如何影响内部受体，内吞机制和微管细胞骨架之间的相互作用，尤其是人类细胞。此外，尚不清楚诸如巨噬细胞等促炎细胞是否对诸如内皮或平滑肌细胞等脆弱靶标（使用Project VI）的响应类似。我们假设循环介质（肾上腺素能，溶酶体和细胞因子）（带有项目V，VII和IX）在创伤增加了膜，受体和随后的信号流量后释放。我们最初专注于为i选择的配体。与创伤有关，ii。 GPCR和III的调解，PKC和MAPK模块的激活。我们将研究这些测试配体如何影响a。构成膜/受体运输和b。来自人类和啮齿动物的一组髓样和非髓样细胞中的内在信号传导。这些研究将构成评估当前可访问的治疗策略是否可以针对网格蛋白涂层的内吞作用和基于微管的贩运的基础。