Force Transduction Mechanisms At Adherens Junctions

机械力是细胞和组织发育的主要决定因素。在组织发育过程中，细胞承受大量的机械力，包括来自周围组织形状变化的拉力或压力，以及来自管状结构中流体流动的剪切力。在肺的衬里中，排列在管状结构上的血管和肾细胞通过连接纤维状内部细胞骨架的连接直接与相邻细胞粘连。电池之间的连接在电池之间传送力。细胞间的连接是肺、肾和血管衬里细胞测量液体流动或气压的力量的一部分。这项研究的目的是模拟流体流动的力是如何耦合到细胞-细胞连接的，以及它们如何导致细胞结构的变化，使细胞正常工作。细胞不能正确感知和传递机械信号会导致肾脏、血管和肺部疾病的进展。由于缺乏工具来测量活细胞中特定蛋白质中的力，我们对力传递途径的了解一直受到限制。这项研究将使用一种新的方法，将基于FRET的力传感器与微流体相结合，从而有机会在流动诱导的细胞骨架力和负责细胞重塑的附着连接(AJ)动力学之间建立联系。特定蛋白质的细胞骨架力的分布及其时间依赖关系将使用FRET探针在受到流动刺激的细胞中进行绘制。贴壁连接动力学将与荧光标记的连接蛋白同时测量。这项研究的结果将是对活细胞中剪切力传递动力学的新理解，也许最重要的是，测量细胞内能量流动的因果关系。FRET测力探头还将为科学界提供一套新的研究工具，用于研究细胞力学。教育的更广泛影响将通过一个独特的实验教育计划或E2E来实现

Mechanical forces are major determinants of cell and tissue development. During tissue development, cells experience numerous mechanical forces including tensile or compressive forces from shape change of the surrounding tissues and shear force from fluid flow in tubular structures. In the lining of the lungs, blood vessels and the kidney cells that line tubular structures adhere directly to their neighbors by junctions that join the fibrous internal cellular cytoskeleton. The cell-to-cell connections transmit forces between the cells. The intercellular connections are part of how lung, kidney and vascular lining cells can measure the forces of fluid flow or air pressure. The objective of this research is to mimic how the forces of fluid flow are coupled to the cell-cell junctions and how they cause cell structural changes in the cells that make them function properly. Failure of cells properly to sense and transmit mechanical signals can cause progression of diseases of the kidney, vasculature and lung.Our understanding of force transmission pathways has been limited due to a lack of tools to measure forces in specific proteins in live cells. This research will use a novel approach of combining FRET based force sensors with microfluidics that presents an opportunity to establish the link between flow-induced cytoskeletal forces and adherens junction (AJ) dynamics responsible for cell remodeling. The distribution of cytoskeletal force and its time-dependence for specific proteins will be mapped using FRET probes in cells subjected to flow stimuli. Adherens junction dynamics will be measured simultaneously with fluorescently labeled junction proteins. The outcome of this research will be a new understanding of the dynamics of shear force transduction in living cells, and perhaps most importantly, a measure of the cause and effect relationships of energy flow in the cell. The FRET force probes would also provide a new research toolset to the scientific community to study cell mechanics. The educational broader impacts will be accomplished through a unique program on Education through Experimentation or E2E