Role of the actin cytoskeleton during respiratory burst in chemoattractant‐stimulated neutrophils

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<jats:title>Abstract</jats:title><jats:p>The aim of this study was to clarify the role of the actin cytoskeleton during chemotactic peptide fMet‐Leu‐Phe (fMLF)‐stimulated respiratory burst in human neutrophil granulocytes. Reactive oxygen species (ROS) was measured as luminol‐amplified chemiluminescence (CL) and F‐actin content as bodipy phallacidin fluorescence in neutrophils treated with latrunculin B or jasplakinolide, an inhibitor and activator of actin polymerization, respectively. Latrunculin B markedly decreased, whereas jasplakinolide increased, the F‐actin content in neutrophils, unstimulated or stimulated with fMLF. Latrunculin B enhanced the fMLF‐triggered ROS‐production more than tenfold. Jasplakinolide initially inhibited the fMLF‐induced CL‐response, however, caused a potent second sustained phase (>400% of control). Both actin drugs triggered a substantial CL‐response when added 5–25 min after fMLF. This was also valid for chemotactic doses of fMLF, where latrunculin B and jasplakinolide amplified the ROS‐production 5–10 times. By using specific signal transduction inhibitors, we found that the NADPH oxidase activation triggered by destabilization of the actin cytoskeleton occurs downstream of phospholipase C and protein kinase C but is mediated by Rho GTPases and tyrosine phosphorylation. In conclusion, rearrangements of the actin cytoskeleton are a prerequisite in connecting ligand/receptor activation, generation of second messengers and assembly of the NADPH oxidase in neutrophil granulocytes.</jats:p>

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