BIOPHYSICAL STUDIES OF CYTOSKELETAL FUNCTION

细胞骨架功能的生物物理学研究

• 批准号: 3295557
• 负责人: Elliot L. Elson
• 金额: $ 21.12万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3295557
• 关键词: Dictyostelium; actins; alpha actinin; biomechanics; cell adhesion; cell cell interaction; cell nucleus; cell type; chromaffin cells; concanavalin A; cytoskeletal proteins; cytoskeleton; extracellular matrix; fibroblasts; fluorescence microscopy; fluorescent dye /probe; human tissue; intermediate filaments; laboratory mouse; leukocytes; membrane activity; myofibrils; myosins; neutrophil; nonblood rheology; pancreatic islet function; physical model; polymerization; secretion; tissue /cell culture; video recording system; viscosity

The long range goal of the proposed work is to understand generally how cells function mechanically and, more specifically, what forces and structures determine their shapes and deformabilities. The approach is based on the hypothesis that cell shape and deformability are governed by the cytoskeleton. Since the cytoskeleton is a system interacting polymer filaments, both its and the cell's viscoelasticity should be understandable in terms of polymer chemistry. To achieve this level of understanding it is necessary to measure quantitatively cellular, and therefore cytoskeletal, viscoelasticity. Toward this end a nev method for measuring cellular viscoelasticity based on determining the resistance of the cell to localized indentation has been developed. This method will be used to determine the relative contributions of the three cytoskeletal systems to the control of cellular deformability by interfering separately with the structure of each of the systems. In addition the mechanical function of the nucleus and of cell-substrate and cell-cell interactions will be examined. The mechanical functions of myosin will be examined in mutant Dictyostelium cells which entirely lack this protein. The mechanical changes resulting from physiological activation of secretory cells and leukocytes will be determined and interpreted in terms of cytoskeletal processes. Supporting this work on cells there will also be studies of the interactions of purified cytoskeletal proteins in simplified reconstituted systems. For example, fluorescence photobleaching recovery will be used to characterize the interactions of alpha-actinin with actin filaments.

拟议工作的长期目标是大致了解如何 细胞的机械功能，更具体地说，是什么力量， 结构决定了它们的形状和可变形性。该方法是 基于细胞形状和变形性受以下因素支配的假设， 细胞骨架由于细胞骨架是一个系统相互作用的聚合物 纤维，它和细胞的粘弹性应该是可以理解的 在聚合物化学方面。为了达到这种理解水平， 必须定量测量细胞，因此细胞骨架， 粘弹性为此，提出了一种新的测量细胞的方法， 粘弹性的基础上确定细胞的阻力， 已经形成了局部压痕。此方法将用于 确定三种细胞骨架系统的相对贡献， 通过单独干扰细胞的变形性来控制细胞的变形性， 每个系统的结构。此外，机械功能 细胞-基质和细胞-细胞相互作用的细胞核将被 考察肌球蛋白的机械功能将在突变体中进行检查。 完全缺乏这种蛋白质的网骨藻细胞。机械 由分泌细胞的生理激活引起的变化， 白细胞将根据细胞骨架来确定和解释 流程.为了支持这项细胞研究，还将对 纯化的细胞骨架蛋白在简化的重组细胞中的相互作用 系统.例如，荧光光漂白恢复将用于 表征α-辅肌动蛋白与肌动蛋白丝的相互作用。