Chemoattractant-stimulated polymorphonuclear leukocytes contain two populations of actin filaments that differ in their spatial distributions and relative stabilities.
Chemoattractants stimulate actin polymerization in lamellipodia of polymorphonuclear leukocytes. We find that removal of chemoattractant results in rapid (within 10 s at 37 degrees C) and selective depolymerization of the F-actin located in lamellipodia. Addition of 10 microM cytochalasin B, in the presence of chemoattractant, also resulted in rapid and selective depolymerization of lamellar F-actin. The elevated F-actin level induced by chemoattractant rapidly returns to the level present in unstimulated cells after (a) a 10-fold decrease in chemoattractant concentration; (b) the addition of 10 microM cytochalasin B; or (c) cooling to 4 degrees C. The F-actin levels of unstimulated cells are only slightly affected by these treatments. Based on the similar effects of cytochalasin addition and chemoattractant dilution, it is likely that both treatments result in actin depolymerization from the pointed ends of filaments. Based on our results we propose that chemoattractant-stimulated polymorphonuclear leukocytes contain two distinct populations of actin filaments. The actin filaments within the lamellipodia are highly labile and in the continued presence of chemoattractant these filaments are rapidly turning over, continually polymerizing at their plus (barbed) ends, and depolymerizing at their minus ends. In contrast, the cortical F-actin filaments of both stimulated and unstimulated cells are differentially stable.
Cassimeris
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Biology Department, University of Pennsylvania, Philadelphia 19104-6018.
McNeill
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Zigmond
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SH
D000199:Actins / Q000032:analysis / Q000097:blood* / Q000648:ultrastructure; D002634:Chemotaxis, Leukocyte; D003571:Cytochalasin B / Q000494:pharmacology; D005456:Fluorescent Dyes; D006801:Humans; D046911:Macromolecular Substances; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D009504:Neutrophils / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure
D000199:Actins; D005456:Fluorescent Dyes; D046911:Macromolecular Substances; D003571:Cytochalasin B