Investigations of the spreading and closure mechanisms of phagocytosis in J774a.1 macrophages

Phagocytosis;Frustrated phagocytosis;Actin;Cytoskeleton;Traction force microscopy;Dynamic contraction model;Structure illumination microscopy;Cell spreading;Cell adhesion

Kovari, Daniel T. ; Curtis, Jennifer E. Physics Fernandez-Nieves, Alberto Goldbart, Paul M. Sulchek, Todd Wiesenfeld, Kurt A. ; Curtis, Jennifer E.

University:Georgia Institute of Technology

Department:Physics

关键词: Phagocytosis;    Frustrated phagocytosis;    Actin;    Cytoskeleton;    Traction force microscopy;    Dynamic contraction model;    Structure illumination microscopy;    Cell spreading;    Cell adhesion;   

Others  :  https://smartech.gatech.edu/bitstream/1853/54882/1/KOVARI-DISSERTATION-2015.pdf

美国|英语

来源: SMARTech Repository

【 摘 要 】

Phagocytosis is the process by which cells engulf foreign bodies. It is the hallmark behavior of white blood cells, being the process through which those cells ingest and degrade pathogens and debris. To date a large amount of research has focused on documenting the existence and role of biochemical components involved with phagocytosis. Scores of signaling molecules have been implicated in the complex signal cascade which drives the process. These molecules are small (typically no larger than 5 nanometers) and operate in a crowded, chemically “noisy,” environment, yet they coordinate the cell's activity over comparatively expansive distances (as large as 20 micrometers). How these molecular processes scale-up to coordinate the activities of the cell over such massive distances is largely unknown. Using a planar analog of phagocytosis termed “frustrated phagocytosis,” we experimentally demonstrate that phagocytosis occurs in three distinct phases: initial cell-antigen binding, symmetric spreading, and late-stage contraction. Initial binding and symmetric spreading appears to be both mechanically and chemically similar to the quasi-universal cellular behaviors of adhesion and migration. Adhesion and migration have received extensive attention from the biophysics community in recent years. Leveraging these similarities, we adapt the biomechanical frameworks used in models of migration to phagocytosis. We show that macroscopic properties such as a cell's effective viscosity and membrane cortical tension can be used to model cell behavior during phagocytosis. Our experiments reveal that late-stage contraction distinguishes frustrated phagocytosis from other spreading behaviors. This contraction is myosin dependent. Additionally we demonstrate, for the first time, that late-stage contraction corresponds with formation of a contractile F-actin belt. Based on the dynamic contraction model (DC) developed to explain actin structure during cell migration we propose a DC model of phagocytosis which posits that contractile belt formation is the result of a late-stage myosin activity coupled with F-actin.