Relationships between phosphoinositide metabolism, Ca2+ changes and respiratory burst in formyl-methionyl-leucyl-phenylalanine-stimulated human neutrophils. The breakdown of phosphoinositides is not involved in the rise of cytosolic free Ca2+.
The relationships between the changes of cellular Ca2+, the activation of phosphoinositide turnover and the functional responses induced by the stimulus-receptor interactions in neutrophils are matter of controversy. By measuring the concentration dependency of different formyl-leucyl-methionyl-phenylalanine (FMLP)-induced changes, the following values of ED50 were found: 1.6 and 0.8 nM for the rise in [Ca2+]i monitored with Quin-2, in the presence and absence of exogenous Ca2+, respectively; 20 nM for the activation of phosphoinositide metabolism, monitored as change in the 32Pi of phosphatidate; 14 nM for membrane-bound Ca2+ mobilization, monitored with chlorotetracycline (CTC); 34 nM for 45Ca2+ influx and 32 nM for the respiratory burst. Furthermore, low dose of FMLP causes an increase in [Ca2+]i in absence of activation of breakdown of phosphatidylinositol, phosphatidylinositol 4-monophosphate and phosphatidylinositol 4,5-biphosphate monitored as changes in [3H]glycerol radioactivity. The results clearly demonstrate that the increase in [Ca2+]i, due to the release from intracellular stores, is not caused by the breakdown of phosphatidylinositides. On the other hand, the data of the similarity of ED50 are compatible with an involvement of phosphoinositide response in the release of membrane bound Ca2+, monitored with CTC, and in the 45Ca influx and in the respiratory burst.
Rossi
F
F
Della Bianca
V
V
Grzeskowiak
M
M
De Togni
P
P
Cabrini
G
G
D000634:Aminoquinolines; D002118:Calcium / Q000097:blood*; D002462:Cell Membrane / Q000378:metabolism; D002751:Chlortetracycline; D005456:Fluorescent Dyes; D006801:Humans; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D009504:Neutrophils / Q000378:metabolism*; D010101:Oxygen Consumption; D010712:Phosphatidic Acids / Q000097:blood; D010716:Phosphatidylinositols / Q000097:blood*
D000634:Aminoquinolines; D005456:Fluorescent Dyes; D010712:Phosphatidic Acids; D010716:Phosphatidylinositols; D009240:N-Formylmethionine Leucyl-Phenylalanine; C035739:Quin2; D002118:Calcium; D002751:Chlortetracycline