A Real Time Chemotaxis Assay Unveils Unique Migratory Profiles amongst Different Primary Murine Macrophages

Chemotaxis assays are an invaluable tool for studying the biological activity of inflammatory mediators such as CC chemokines, which have been implicated in a wide range of chronic inflammatory diseases. Conventional chemotaxis systems such as the modified Boyden chamber are limited in terms of the...

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Vydáno v:	PloS one Ročník 8; číslo 3; s. e58744
Hlavní autoři:	Iqbal, Asif J., Regan-Komito, Daniel, Christou, Ivy, White, Gemma E., McNeill, Eileen, Kenyon, Amy, Taylor, Lewis, Kapellos, Theodore S., Fisher, Edward A., Channon, Keith M., Greaves, David R.
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	United States Public Library of Science 14.03.2013 Public Library of Science (PLoS)
Témata:	Actin Analysis Animals Antigens Assaying Biological activity Biology Bone marrow Bone Marrow Cells - cytology Boyden chamber CC chemokines CD14 antigen Cell migration Chemokines Chemokines, CC - pharmacology Chemokinesis Chemotaxis Chemotaxis - drug effects Cytochalasin D Cytokines Cytoskeleton Cytoskeleton - drug effects Cytoskeleton - metabolism Ethics Gene expression GTP-Binding Protein alpha Subunits, Gi-Go - metabolism Humans Inflammation Inflammatory diseases Laboratories Macrophages Macrophages - cytology Male Medicine Mice Mice, Inbred C57BL Monocyte chemoattractant protein 1 Monocytes Monocytes - cytology Monocytes - drug effects Pathology Penicillin Pertussis Pertussis toxin Populations Proteins Real time Signal transduction Signal Transduction - drug effects Signaling Time Factors Toxins New York United States > US
ISSN:	1932-6203, 1932-6203
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Shrnutí:	Chemotaxis assays are an invaluable tool for studying the biological activity of inflammatory mediators such as CC chemokines, which have been implicated in a wide range of chronic inflammatory diseases. Conventional chemotaxis systems such as the modified Boyden chamber are limited in terms of the data captured given that the assays are analysed at a single time-point. We report the optimisation and validation of a label-free, real-time cell migration assay based on electrical cell impedance to measure chemotaxis of different primary murine macrophage populations in response to a range of CC chemokines and other chemoattractant signalling molecules. We clearly demonstrate key differences in the migratory behavior of different murine macrophage populations and show that this dynamic system measures true macrophage chemotaxis rather than chemokinesis or fugetaxis. We highlight an absolute requirement for Gαi signaling and actin cytoskeletal rearrangement as demonstrated by Pertussis toxin and cytochalasin D inhibition. We also studied the chemotaxis of CD14(+) human monocytes and demonstrate distinct chemotactic profiles amongst different monocyte donors to CCL2. This real-time chemotaxis assay will allow a detailed analysis of factors that regulate macrophage responses to chemoattractant cytokines and inflammatory mediators.
Bibliografie:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 Competing Interests: The authors have declared that no competing interests exist. Conceived and designed the experiments: AJI GEW EAF KMC DRG. Performed the experiments: AJI DRK IC EM AK LT TSK DRG. Analyzed the data: AJI IC DRG. Contributed reagents/materials/analysis tools: AJI GEW EM KMC DRG. Wrote the paper: AJI GEW EAF DRG.
ISSN:	1932-6203 1932-6203
DOI:	10.1371/journal.pone.0058744