title: Analysis of neutrophil polarization and chemotaxis.
abstract: Neutrophil polarization and directed migration (chemotaxis) are critical for the inflammatory response. Neutrophil chemotaxis is achieved by the sensing of narrow gradients of chemoattractant and the subsequent polarization and directed migration toward the chemotactic source. Despite recent progress, the signaling mechanisms that regulate neutrophil polarization during chemotaxis have not been clearly defined. Here, we describe methods to analyze neutrophil polarization and asymmetric redistribution of signaling components induced by chemoattractant using immunofluorescence. Further, methods are described to dissect the role of specific signaling pathways during chemotaxis by the use of murine neutrophils from transgenic mouse models. Finally, methods for time-lapse microscopy and transwell assay for the analysis of neutrophil chemotaxis will also be discussed.
authors.affiliation: Department of Pediatrics, University of Wisconsin Medical School, Madison, WI, USA.
authors.forename: Mary A
authors.initials: MA
authors.lastname: Lokuta
authors.forename: Paul A
authors.initials: PA
authors.lastname: Nuzzi
authors.forename: Anna
authors.initials: A
authors.lastname: Huttenlocher
mesh_terms: D000818:Animals; D001781:Blood Component Removal; D002469:Cell Separation; D002634:Chemotaxis, Leukocyte / Q000502:physiology*; D005404:Fixatives / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D009504:Neutrophils / Q000502:physiology*; D013194:Staining and Labeling
chemical_list: D005404:Fixatives