Microscopic co-distributions of myosin, actin, alpha-actinin and vinculin in human neutrophils during movement.
Human neutrophils initiate their characteristic ameboid movement in response to external stimuli via a mechanochemical system of contractile proteins using mechanical energy derived from hydrolysis of adenosine triphosphate. We have visualized contractile proteins, including myosin, filamentous actin (F-actin), alpha-actinin, tropomyosin, and vinculin in human neutrophils during movement by fluorescence microscopy using double immunofluorescence staining procedures of fluorescein isothiocyanate-labeled antibodies, rhodamine-labeled antibodies, or rhodamine-conjugated phalloidin. From dual color images, both myosin and F-actin, both alpha-actinin and F-actin, both vinculin and F-actin showed co-localization, both myosin and alpha-actinin showed a partial co-localization, at the leading edge in the lamellipodia, and both vinculin and alpha-actinin were co-localized at the leading edge and appeared reciprocally at regular intervals in the lamellipodia. Our observations indicate that most of myosin, F-actin, alpha-actinin, and vinculin are focused in order to reorganize dynamically at the leading edge in the lamellipodia, and a motile apparatus is reconstituted there, when neutrophils start to move toward external substances.
Takubo
T
T
Department of Clinical and Laboratory Medicine, Osaka City University Medical School, Osaka, Japan. m3542254@med.osaka-u.ac.jp
Tatsumi
N
N
D000185:Actinin / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D000328:Adult; D002465:Cell Movement; D003285:Contractile Proteins / Q000378:metabolism*; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods; D009218:Myosins / Q000378:metabolism; D009504:Neutrophils / Q000166:cytology* / Q000276:immunology; D014018:Tissue Distribution; D016596:Vinculin / Q000378:metabolism*
D000199:Actins; D003285:Contractile Proteins; D000185:Actinin; D016596:Vinculin; D009218:Myosins