Motility of human polymorphonuclear neutrophils: Microscopic analysis of substrate adhesion and distribution of F-actin

摘要: Directed movement of polymorphonuclear neutrophils (PMN) requires cell polarization and the orderly making breaking cell-substrate contacts. We compared human PMN suspended from underside glass coverslips to that seen in “profile” on fibers, using brightfield, differential interference contrast reflection microscopy. Images were recorded film videotape analyzed real time lapse. The distribution F-actin was observed with image-enhanced fluorescence microscopy after staining NBD-phallacidin. PMN exhibited two patterns motility. Fifteen twenty-five percent cells moved a low profile gliding pattern cauded displacement dorsal surface folds. Most made progress by cycles partial release lamellipodium substrate anterior advance followed arching or rolling lamellipodial reassociation substrate. Cells stimulated bacteria, casein, chemotactic formyl peptide rarely spread coverglass but waved into medium attached only uropod. Eventually, many detached completely confined overlying agarose able locomote when confronted these substances. F-actin irregularly distributed nonpolarized concentrated polarized cells. As arched along substrate, accumulated foci corresponding substrate-PMN interface, particularly at uropod retraction fibrils. Conversely, physically restricted plane diffuse entire cell. Suspended incubated Con A These observations suggest both locomotion binding sites involve caudad redistribution F-actin.