Chemoattractant-induced cytoplasmic pH changes and cytoskeletal reorganization in human neutrophils. Relationship to the stimulated calcium transients and oxidative burst.
The relationships between the chemotactic factor-stimulated mobilization of calcium, activation of the NADPH-oxidase, changes in cytosolic pH, and in the level of polymerized actin in human neutrophils have been examined. The approach taken was to use intracellular calcium chelators, and pharmacologic modulators (both positive and negative) of the NADPH-oxidase to measure the aforementioned responses under conditions where the calcium transients were abrogated and/or the generation of superoxide anions was either inhibited or augmented. The decrease in cytosolic pH induced by chemoattractants was inhibited by the calcium chelator BAPTA and by the diglyceride kinase inhibitor 6-[2-(4-[(4-fluorophenyl)phenylmethylene]-1-piperidinylethyl ]-7-methyl-5H-thiazolo[3,2-alpha]pyriimidin-5-one (R59022) (this latter compound enhanced the oxidative response of the cells). Furthermore, a specific inhibitor of the NADPH-oxidase (diphenyleneiodonium) had no significant effect on the cytosolic acidification induced by FMLP or leukotriene B4. These results indicate that the initiation of the cytosolic acidification induced by chemotactic factors is a calcium-dependent event that is not directly linked to the activation of the NADPH-oxidase. In contrast, the stimulated polymerization of actin was insensitive to BAPTA, R59022, and diphenyleneiodonium. Thus, neither the calcium transients nor the oxidative burst play a signaling role in the initiation of actin polymerization elicited by chemoattractants. These data indicate that additional investigations are needed to uncover the biochemical basis of the signals initiated in human neutrophils by chemotactic factors that lead to the polymerization of actin and to the cytosolic acidification.
Naccache
P H
PH
Département de Médecine, Université Laval, Ste Foy, Québec, Canada.
Therrien
S
S
Caon
A C
AC
Liao
N
N
Gilbert
C
C
McColl
S R
SR
D000199:Actins / Q000378:metabolism / Q000502:physiology*; D001667:Binding, Competitive; D000001:Calcimycin; D002118:Calcium / Q000502:physiology*; D002630:Chemotactic Factors; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D004533:Egtazic Acid / Q000494:pharmacology / Q000031:analogs & derivatives; D006801:Humans; D007975:Leukotriene B4; D046911:Macromolecular Substances; D009240:N-Formylmethionine Leucyl-Phenylalanine; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D009861:Onium Compounds; D010101:Oxygen Consumption / Q000187:drug effects; D011744:Pyrimidinones; D013481:Superoxides / Q000096:biosynthesis; D013844:Thiazoles
D000199:Actins; D002630:Chemotactic Factors; D046911:Macromolecular Substances; D009861:Onium Compounds; D011744:Pyrimidinones; D013844:Thiazoles; D013481:Superoxides; D007975:Leukotriene B4; D000001:Calcimycin; D004533:Egtazic Acid; D009240:N-Formylmethionine Leucyl-Phenylalanine; C007517:diphenyleneiodonium; C047505:R 59022; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D002118:Calcium