Piezo1 Mobility Dynamics in Mechanotransduction

机械传导中的 Piezo1 移动动力学

• 批准号: 10612807
• 负责人: Alan Ly
• 金额: $ 4.25万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10612807
• 关键词: Acids; Actins; Actomyosin; Affect; Agonist; Architecture; Area; Behavior; Benzyl Alcohols; Biological Process; Biophysics; Blood Vessels; Cell Polarity; Cell Shape; Cell membrane; Cells; Characteristics; Chemicals; Cholesterol; Cytoskeletal Modeling; Cytoskeleton; Data; Defect; Development; Diffusion; Embryo; Environment; Fibroblasts; Fluorescence Microscopy; Goals; Human; Image; Imaging Techniques; Ion Channel; Ion Channel Gating; Label; Link; Lipids; Measurement; Mechanics; Membrane; Membrane Proteins; Molecular; Mus; Neurodegenerative Disorders; Neurodevelopmental Disorder; Pattern; Physiological; Physiological Processes; Piezo 1 ion channel; Play; Process; Proliferating; Protein Dynamics; Reporting; Research; Role; Shapes; Signal Transduction; Skeletal Muscle; System; Testing; Therapeutic; Time; Traction; Work; antagonist; blood pressure regulation; body system; cell type; experience; experimental study; gain of function mutation; genetic manipulation; human imaging; induced pluripotent stem cell; inhibitor; insight; ion dynamics; mechanical force; mechanical signal; mechanotransduction; migration; nerve stem cell; neural repair; neurodevelopment; non-invasive imaging; novel; particle; pharmacologic; polarized cell; spatiotemporal; stem cell fate; transduction efficiency

Project Summary/Abstract: The mechanically-activated ion channel Piezo1 plays diverse roles in various physiological processes, including the differentiation of neural stem/progenitor cells. However, the subcellular diffusion dynamics of the channel, and how these characteristics contribute to its function, are unknown. Single-particle tracking analysis suggests that Piezo1 diffusion is affected by cellular architecture (e.g. actin cytoskeleton and the lipid environment). We have previously reported in square-shaped cells that Piezo1 is most active in high traction force regions (edges and vertices). Given Piezo1’s diverse role in mechanotransduction and neurodevelopment, it is necessary to understand the role of Piezo1 diffusion in these biological processes. I hypothesize that channel activity and cellular mechanics determine Piezo1 diffusion dynamics. I will perform single-particle tracking measurements of endogenous Piezo1 channels in human induced pluripotent stem cell-derived neural stem cells (hiPSC-NSCs) via Total Internal Reflection Fluorescence microscopy (TIRFM). I will image and analyze Piezo1 diffusion in conditions where (1) Piezo1 activity is pharmacologically and genetically manipulated and (ii) cellular mechanics is manipulated . The proposed research will elucidate the function of Piezo1’s diffusion in mechanotransduction and bridge the gap between Piezo1 activity and diffusion.

项目概要/摘要： 机械激活的离子通道Piezo 1在各种生理过程中起着不同的作用， 包括神经干/祖细胞的分化。然而，亚细胞扩散动力学的 通道，以及这些特征如何有助于其功能，是未知的。 单粒子跟踪分析表明，Piezo 1扩散受细胞结构（例如， 肌动蛋白细胞骨架和脂质环境）。我们以前曾报道，在方形细胞，Piezo 1 在高牵引力区域（边缘和顶点）中最活跃。鉴于Piezo 1在其中的不同角色 机械转导和神经发育，有必要了解Piezo 1扩散在 这些生物过程。 我假设通道活动和细胞力学决定Piezo 1扩散动力学。我会 进行人诱导多能干细胞中内源性Piezo 1通道的单粒子跟踪测量 干细胞衍生的神经干细胞（hiPSC-NSC）通过全内反射荧光显微镜 （TIRFM）。我将在以下条件下对Piezo 1扩散进行成像和分析：（1）Piezo 1活性降低 和基因操纵和（ii）细胞力学被操纵。 拟议的研究将阐明Piezo 1扩散在机械传导中的作用， 弥合压电活性和扩散之间的差距。