Actin polymerization in neutrophils is triggered without a requirement for a rise in cytoplasmic Ca2+.
Stimulation of rat neutrophils with the peptide fMetLeuPhe caused (i) the appearance of a 40 kDa protein in the Triton-X-100-insoluble cytoskeleton, (ii) the disappearance of DNAase inhibition from the cytosol and (iii) the appearance of N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)phallacidin (NBD-phallacidin) binding sites. All three observations were consistent with a rapid and transient assembly of polymerized actin, peaking at approximately 5 s and returning to near resting levels within 40 s. By experimentally depleting the cells of Ca2+ and increasing the cytoplasmic Ca2+ buffering capacity, the peptide-induced Ca2+ transient was reduced from a peak of 900 nM to 250 nM, without inhibiting actin polymerization, and this peak was sustained for at least 2 min. A further dissociation between the triggering of actin polymerization and peptide-induced Ca2+ elevation and oxidase activation was demonstrated at low concentrations of peptide (1-100 pM), actin polymerization being triggered without an elevation in Ca2+ or activation of the oxidase. Two other agents which induced actin polymerization, phorbol 12-myristate 13-acetate and latex beads, failed to elevate cytoplasmic Ca2+. It was therefore concluded that neither Ca2+ nor those intracellular messengers which act with Ca2+ to trigger the neutrophil oxidase are responsible for triggering actin polymerization in neutrophils.
al-Mohanna
F A
FA
Department of Surgery, University of Wales College of Medicine, Cardiff, U.K.
Hallett
M B
MB
D000199:Actins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000546:Amanitins; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000494:pharmacology; D005089:Exocytosis; D008970:Molecular Weight; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism; D010587:Phagocytosis; D051381:Rats; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic / Q000502:physiology
D000199:Actins; D000546:Amanitins; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic; C028877:7-nitrobenz-2-oxa-1,3-diazole-phallacidin; D010088:Oxidoreductases; D003850:Deoxyribonuclease I; D002118:Calcium