Analysis of Physical Mechanisms in Cell Adhesion

描述(由申请人提供)： 控制炎症是提高工程组织和移植组织生存能力的关键。考虑到这一点，我们的工作阐明了促进中性粒细胞募集到炎症组织的膜重塑事件。第一个目的是利用光漂白后荧光恢复(FRAP)、全内反射荧光(TIRF)和图像互相关光谱(ICCS)来研究黏附调节的物理机制，包括整合素和选择素受体的迁移率和定位。这些工具的互补性将以高于以往所有工作的信心和准确性来评估受体的位置和移动性。将这些技术与药物治疗相结合，第二个目的是检验一个假设，即由于短暂的细胞骨架释放，黏附分子在激活的中性粒细胞上随着侧向迁移率的增加而重新分布。第三个目的是研究迁移性中性粒细胞。利用定量显微镜和生化处理可以同时显示表面受体和细胞骨架，这一目的假设高整合素迁移率和低选择素迁移率是迁移的固有特征，并与潜在的细胞骨架结构相关。

DESCRIPTION (provided by applicant): Controlling inflammation is key to improving the viability of engineering and transplanted tissues. With this in mind, our work elucidates the membrane remodeling events that facilitate recruitment of neutrophils to inflamed tissues. The first aim examines the physical mechanisms of adhesion regulation including mobility and localization of integrin and selectin receptors using fluorescence recovery after photobleaching (FRAP), total internal reflection fluorescence (TIRF), and image cross-correlation spectroscopy (ICCS). The complementary nature of these tools will assess the position and mobility of receptors with superior confidence and accuracy over all previous work. Employing these techniques with pharmacological treatments, the second aim tests a hypothesis that adhesion molecules redistribute with increased lateral mobility on activated neutrophils because of transient cytoskeletal release. The third aim examines migratory neutrophils. Using quantitative microscopy and biochemical processing that allows simultaneous visualization of surface receptors and cytoskeleton, this aim hypothesizes that high integrin mobility and low selectin mobility are inherent features of migration and correlated with underlying cytoskeletal structure.