Analysis of Physical Mechanisms in Cell Adhesion

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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.