Cell mechanics and mechanical transduction by ion channels

细胞力学和离子通道的机械转导

• 批准号: 7741790
• 负责人: FREDERICK SACHS
• 金额: $ 72.51万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7741790
• 关键词: Accounting; Address; Animals; Arrhythmia; Biology; Bulla; Cell Line; Cell membrane; Cells; Cellular Stress; Cholesterol; Cytoskeletal Proteins; Cytoskeleton; Data; Disease; Edema; Electric Capacitance; Electrophysiology (science); Environment; Extracellular Matrix; Family; Fluorescent Probes; Glaucoma; Hearing; Heat shock proteins; Hypertension; Ion Channel; Ion Channel Gating; Kinetics; Label; Link; Lipid Bilayers; Maps; Measurement; Measures; Mechanical Stress; Mechanics; Membrane; Microscopy; Movement; Muscle Contraction; Muscular Dystrophies; Optics; Pathology; Physiology; Preparation; Process; Property; Proteins; Relaxation; Resolution; Scanning; Signal Transduction; Stress; TimeLine; Touch sensation; Work; base; blood pressure regulation; cell behavior; cell cortex; deafness; fluorescence imaging; hemodynamics; member; patch clamp; potassium channel protein TREK-1; pressure; protein expression; public health relevance; research study; scanning ion conductance microscopy; stress protein

DESCRIPTION (provided by applicant): Animal cell behavior is tightly linked to mechanical stresses produced by the environment and mechanochemistry illustrated most vividly by muscle contraction and hearing. There is an enormous amount of physiology and disease connected to forces and sensing including hemodynamics, coordinated movement, touch, cardiac arrhythmias, muscular dystrophy, edema, glaucoma, deafness, high blood pressure, etc. The cell cortex forms the interface between the environment and the cell and this project addresses the distribution of stress in the cell cortex and how it is sensed by mechanosensitive ion channels (MSCs). We will analyze how stress is shared by specific cytoskeletal proteins, the lipid bilayer, and sensed by mechanosensitive ion channels. The project has two specific aims directed at patches and whole cells and creating a basis for extrapolating from high resolution patch data to cell behavior. The patch experiments will localize different channels and cytoskeletal proteins within the patch; measure stress in cytoskeletal proteins as the patch is stressed; create patches with minimal cytoskeleton to simplify the stress distribution; measure and quantify endogenous and TREK-1 channel kinetics in terms of membrane tension; characterize the properties of microdomains within the patch using channel kinetics, patch capacitance, fluorescence imaging of labeled proteins and the stress in specific labeled cytoskeletal proteins. The whole cell experiments will use a combination of scanning conductance microscopy (SICM), whole cell patch clamp and fluorescent probes to examine the distribution of cytoskeletal proteins and channels and the stress in cytoskeletal proteins as the cell are stressed by the SICM. We will use MSCs calibrated in the patch as additional probes of bilayer stress. PUBLIC HEALTH RELEVANCE: Mechanical forces in biology account for hearing, muscle contraction, blood pressure regulation and a great deal more. As expected for such ubiquitous processes, they are also involved in much pathology such as cardiac arrhythmias, high blood pressure and muscular dystrophy. This proposal analyses how stresses are distributed in molecules, membranes and cells and how that leads to signal transduction by ion channels.

描述（由申请人提供）：动物细胞行为与环境和机械化学产生的机械应力紧密相关，肌肉收缩和听觉最生动地说明了这一点。大量的生理学和疾病与力和传感相关，包括血流动力学、协调运动、触觉、心律失常、肌营养不良、水肿、青光眼、耳聋、高血压等。细胞皮层形成环境和细胞之间的界面，该项目解决了细胞皮层中压力的分布以及机械敏感离子通道 (MSC) 如何感知压力。我们将分析特定细胞骨架蛋白、脂质双层如何共享压力以及如何通过机械敏感离子通道感知压力。该项目有两个具体目标，针对斑块和整个细胞，并为从高分辨率斑块数据推断细胞行为奠定基础。贴片实验将定位贴片内的不同通道和细胞骨架蛋白；当贴片受到压力时，测量细胞骨架蛋白的压力；创建具有最小细胞骨架的补丁以简化应力分布；根据膜张力测量和量化内源性和 TREK-1 通道动力学；使用通道动力学、贴片电容、标记蛋白的荧光成像以及特定标记的细胞骨架蛋白中的应力来表征贴片内微域的特性。全细胞实验将使用扫描电导显微镜 (SICM)、全细胞膜片钳和荧光探针的组合来检查细胞骨架蛋白和通道的分布以及当细胞受到 SICM 应激时细胞骨架蛋白的应激。我们将使用贴片中校准的 MSC 作为双层应力的附加探针。公共健康相关性：生物学中的机械力负责听力、肌肉收缩、血压调节等。正如对这种普遍存在的过程所预期的那样，它们还涉及许多病理学，例如心律失常、高血压和肌营养不良症。该提案分析了压力如何在分子、膜和细胞中分布，以及如何导致离子通道的信号转导。